The transformation from ductal carcinoma in situ (DCIS) to invasive breast cancer (IBC) is a crucial step in breast cancer progression. The specific alterations that govern this transition have not been elucidated. HER2/neu is frequently overexpressed in DCIS but is less common in IBC suggesting additional requirements for transformation. To identify genes capable of cooperating with HER2/neu to fully transform mammary epithelial cells, we used an insertional mutagenesis screen on cells isolated from wild-type neu expressing mice and identified the E3 ligase HACE1 as a HER2 cooperative tumor suppressor gene. Loss of HACE1 expression is commonly seen in clinical multiple cancer datasets including breast cancer. HACE1 down-regulation in normal human mammary epithelial cells (HMECs) results in the accumulation of the activated GTP-bound Rac1 partially transforming these cells. Overexpression of HER2 activates Rac1, which further accumulates upon HACE1 loss resulting in Rac1 hyperactivation. While the
Define insertional mutagenesis. insertional mutagenesis synonyms, insertional mutagenesis pronunciation, insertional mutagenesis translation, English dictionary definition of insertional mutagenesis. Noun 1. insertional mutagenesis - a mutation caused by the insertion of exogenous DNA into a genome genetic science, genetics - the branch of biology that...
Fast MultiSite Mutagenesis System,Mutagenesis System,Cloning and Mutagenesis System,Products,Beijing TransGen Biotech Co.Ltd,OverviewContents& storageCitations & referencesRelated ImagesDownloadOverviewDescriptionFast MultiSite Mutagenesis Syst
TY - JOUR. T1 - Cc.snf5, a gene encoding a putative component of the SWI/SNF chromatin remodeling complex, is essential for sexual development in the agaricomycete Coprinopsis cinerea. AU - Ando, Yuki. AU - Nakazawa, Takehito. AU - Oka, Kunihiko. AU - Nakahori, Kiyoshi. AU - Kamada, Takashi. PY - 2013/1. Y1 - 2013/1. N2 - We characterized a Coprinopsis cinerea mutant strain, Spe20, defective in fruiting initiation, which was isolated after restriction enzyme-mediated integration (REMI) mutagenesis of a homokaryotic fruiting strain, 326. A plasmid rescue followed by complementation experiments, RACE, and cDNA analyses revealed that the gene, a mutation of which is responsible for the phenotype, is predicted to encode a protein that exhibits a high similarity to yeast Snf5p, a key component of the chromatin remodeling complex SWI/SNF, and named Cc.snf5. Cc.Snf5 is, however, different from Snf5p in that the former has, in addition to an Snf5 domain comprising N-terminal repeat1 (rp1) and C-terminal ...
Tn-Seq is an experimental method for probing the functions of genes through construction of complex random transposon insertion libraries and quantification of each mutants abundance using next-generation sequencing. An important emerging application of Tn-Seq is for identifying genetic interactions, which involves comparing Tn mutant libraries generated in different genetic backgrounds (e.g. wild-type strain versus knockout strain). Several analytical methods have been proposed for analyzing Tn-Seq data to identify genetic interactions, including estimating relative fitness ratios and fitting a generalized linear model. However, these have limitations which necessitate an improved approach. We present a hierarchical Bayesian method for identifying genetic interactions through quantifying the statistical significance of changes in enrichment. The analysis involves a four-way comparison of insertion counts across datasets to identify transposon mutants that differentially affect bacterial fitness
Insertional mutagenesis approaches use oncoretroviruses or transposons to trigger cancer in mice by widespread integration into the cellular genome and activation of oncogenes near the integration site. Mapping the genomic integration sites in tumors allows the identification of genomic regions that are recurrently hit in independent tumors (defined as Common Insertion Sites, CIS) (5), which host genes likely involved in cancer development (6).. We have recently developed a new specifically tailored LV-based insertional mutagen that allowed the induction of hepatocellular carcinoma in 3 different mouse models (4). In order to identify the integration sites from LV-induced tumors and control nontumoral samples, we exploited LAM-PCR (1) followed by 454 pyrosequencing. The analysis of LV integrations in tumors allowed the identification of 4 new liver cancer genes. Here we describe how we applied the LAM-PCR protocol1 for the retrieval of LV integrations from the hepatocellular carcinomas induced ...
Read "Strategies for Mutagenesis and Gene Cloning Using Transposon Tagging and T-DNA Insertional Mutagenesis, Annual Review of Plant Biology" on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips.
The IGTC focuses on creating resources of embryonic stem cells with gene trap insertions in every or most genes in the mouse genome. A brief description of the gene trap consortium is available online. The Gene Trap Consortium is also described in Nature Genetics 36: 543-544, 2004. Links to Databases of Gene Trapped ES Cell Lines:. ...
Systems and methods for isolating DNA molecules that can include the steps of: providing a transposon mutant collection, the transposon mutant collection being stored in a plurality of wells; dispatc
OFDM (orthogonal frequency division multiplexing) communication is very attractive for high data rate transmission especially in the frequency selective fa
A novel P[UAS] insertion line shows progressive behavioral defects.Crossing P[UAS]117 to the pdf-gal4 driver results in a significant decrease in the rhythmicit
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Insertion Verification by PCR:. We will make fly DNA from transgenic stocks and perform PCR to confirm insertion at the targeted site. $25 per line.. Virgin Collection:. ...
An insertion is an addition in which an atom in one reactant is inserted between two atoms bound to each other by a covalent bond in the other reactant.. ...
... was invented by Researchers at the Johns Hopkins High Throughput Biology Center. Tip-chip can be used to help identify otherwise elusive disease-causing mutations in the 97 percent of the genome long believed to be
Promoterless gene trap vectors have been widely used for high-efficiency gene targeting and random mutagenesis in embryonic stem (ES) cells. Unfortunately, such vectors are only effective for genes expressed in ES cells and this has prompted the development of expression-independent vectors. These polyadenylation (poly A) trap vectors employ a splice donor to capture an endogenous genes polyadenylation sequence and provide transcript stability. However, the spectrum of mutations generated by these vectors appears largely restricted to the last intron of target loci due to nonsense-mediated mRNA decay (NMD) making them unsuitable for gene targeting applications. Here, we present novel poly A trap vectors that overcome the effect of NMD and also employ RNA instability sequences to improve splicing efficiency. The set of random insertions generated with these vectors show a significantly reduced insertional bias and the vectors can be targeted directly to a 5′ intron. We also show that this ...
Looking for online definition of interrupted gene in the Medical Dictionary? interrupted gene explanation free. What is interrupted gene? Meaning of interrupted gene medical term. What does interrupted gene mean?
We are attempting to identify cellular oncogenes activated in mammary tumours by using the mouse mammary tumour virus (MMTV) as an insertional mutagen. MMTV, a retrovirus lacking a host cell-derived viral oncogene, induces adenocarcinomas of the mammary gland after a long latency period. The tumours are clonal outgrowths of cells carrying one or more integrated MMTV proviral copies. We have cloned an integrated MMTV provirus with its adjacent chromosomal DNA and we have established that the insertion site was part of a domain of the mouse genome in which MMTV proviruses are inserted in many different tumours. A gene within this domain, called int-1 is transcriptionally activated as a consequence of proviral integration. We have proposed that int-1 is a cellular oncogene for mammary tumours. Proviral activation of int-1 occurs in cis, over distances of up to 10 kilobases and is presumably caused by the transcriptional enhancer present on the MMTV long terminal repeat. The putative int-1 mammary ...
The PiggyBac (PB) transposon has emerged as a novel mutagenesis tool for understanding gene function and for phenotypic screening in eukaryotes
Multiple methods have been introduced over the past 30 years to identify the genomic insertion sites of transposable elements and other DNA elements that integrate into genomes. However, each of these methods suffer from limitations that can frustrate attempts to map multiple insertions in a single genome and to map insertions in genomes of high complexity that contain extensive repetitive DNA. I introduce a new method for transposon mapping that is simple to perform, can accurately map multiple insertions per genome, and generates long sequence reads that facilitate mapping to complex genomes. The method, called TagMap, for Tagmentation-based Mapping, relies on a modified Tn5 tagmentation protocol with a single tagmentation adaptor followed by PCR using primers specific to the tranposable element and the adaptor sequence. Several minor modifications to normal tagmentation reagents and protocols allow easy and rapid preparation of TagMap libraries. Short read sequencing starting from the adaptor ...
Transposon mutagenesis is one of the most widely used strategies to generate a large number of random mutations within a bacterial genome and then to precisely ...
Transposons have been used in invertebrates for transgenesis and insertional mutagens in genetic screens. We tested a functional transposon called Sleeping Beauty in the one-cell mouse embryo. In this report, we describe experiments in which transposon vectors were injected into one-cell mouse embryos with mRNA expressing the SB10 transposase enzyme. Molecular evidence of transposition was obtained by cloning of insertion sites from multiple transgenic mice produced by SB10 mRNA/transposon coinjection. We also demonstrate germ-line transmission and expression from transposed elements. This technique has promise as a germ-line transgenesis method in other vertebrate species and for insertional mutagenesis in the mouse ...
An alternative to insulin injections is the insulin pump. The pump is a computerized device, about the size of a beeper or pager, often worn on a belt or in a pocket. The pump delivers a continuous low (basal) dose of insulin through a cannula (a flexible plastic tube), which attaches to the body through a small needle inserted into the skin. The cannula is taped in place and the needle is removed. Common insertion sites on the body include the thighs, buttocks, abdomen, upper arms and other areas with fatty tissue.. When a person wearing a pump eats, she pushes a button on the pump to deliver an extra amount of insulin called a bolus to provide insulin for their food.. The advantages of the pump include:. ...
An ampicillin resistance plasmid carrying the cloned repressor gene cII of the L phage (Salmonella lyphimurium) was conducted by Flac into an F- recipient. Two types of plaamids were isolated from Apr transconjugants. The majority of plasmids were dimers with one copy of Tn1000 inserted, the minority being monomers with one copy of Tn1000. This proportion remained unaltered when we used the Flac strain transformed with a monomeric form of the recombinant plasmid as a donor. An extensive oligomerization of pBR322-originating plasmids was proved in the presence of Flac; its presumable relationship to transposition-related processes is suggested.
Transposon insertion in ykyB increases the activation of an artificial ComK feedback loop.Strains PG401 (amyE::PcomG-lacZ-gfp, PcomG-comK, ΔmecA) and PG401-Tn4
An important aspect of functional genomics is understanding the phenotypic consequence of mutations in all genes in a genome. A comprehensive collection of gene knockouts allows a defined set of mutations to be systematically studied for more efficient association of genes to functions (reviewed in [1]). Multiple approaches have been used to develop comprehensive knockout resources. Biological differences between organisms make specific technologies such as homologous recombination, RNA interference, or insertional mutagenesis more useful in generating a resource for an individual species. In plants, a comprehensive knockout collection was generated for Arabidopsis thaliana via mutagenesis with insertion tags [2-5]. Genomic DNA flanking each tag was systematically amplified and sequenced from each mutant. These Flanking Sequence Tags (FSTs) index each mutant to the genome and are accessible through the SIGnAL T-DNA Express database, which links the mutant stocks to genome annotations [6]. A ...
에탄올, 고온, 저해제 스트레스 상황에서 내성을 가지는 사카로마이세스 세레비지아에 균주는 에탄올 생산 산업을 위해 중요한 부분을 차지한다. 본 연구에서는 에탄올 15 % 까지 내성을 가지는 5개의 균주를 트랜스포존에 의해 돌연변이가 일어난 집단에서 테스트를 통해 분리했다. 5개중에 2개는 고온 (42 °C)에도 내성을 가지는 것을 조사되었고, 그 중에서 1개는 푸르푸랄 50 mM까지 내성을 가지는 것을 확인했다. 트랜스포존이 끼워 들어간 위치를 통해 7개의 추정 유전자 (CMP2, IMD4, SSK2, PPG1, DLD3, PAM1, MSN2)를 확인하고, 노던 블럿 분석법을 통해 CMP2와 IMD4, SSK2와 PPG1유전자가 동시에 하향조절 되었고, DLD3 유전자또한 하향조절, PAM1과 MSN2의 전사 해독 틀이 깨져있는 것을 확인 할 수 있었다. 이 7 유전자가 독립적으로 망가진 돌연변이는 에탄올 내성이 있었고, 그들 ...
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TY - JOUR. T1 - Insertional mutagenesis and cloning of the genes required for biosynthesis of the host-specific AK-toxin in the Japanese pear pathotype of Alternaria alternata. AU - Tanaka, Aiko. AU - Shiotani, Hiroshi. AU - Yamamoto, Mikihiro. AU - Tsuge, Takashi. PY - 1999/8. Y1 - 1999/8. N2 - The Japanese pear pathotype of Alternaria alternata causes black spot of Japanese pear by producing a host-specific toxin known as AK-toxin. Restriction enzyme-mediated integration (REMI) mutagenesis was used to tag genes required for toxin biosynthesis. Protoplasts of a wild-type strain were treated with a linearized plasmid along with the restriction enzyme used to linearize the plasmid. Of 984 REMI transformants recovered, three produced no detectable AK-toxin and lost pathogenicity on pear leaves. Genomic DNA flanking the integrated plasmid was recovered from one of the mutants. With the recovered DNA used as a probe, a cosmid clone of the wild-type strain was isolated. Structural and functional ...
Transposons are mobile DNA segments that can disrupt gene function by inserting in or near genes. Here, we show that insertional mutagenesis by the PiggyBac transposon can be used for cancer gene discovery in mice. PiggyBac transposition in genetically engineered transposon-transposase mice induced cancers whose type (hematopoietic versus solid) and latency were dependent on the regulatory elements introduced into transposons. Analysis of 63 hematopoietic tumors revealed that PiggyBac is capable of genome-wide mutagenesis. The Piggybac screen uncovered many cancer genes not identified in previous retroviral or Sleeping Beauty transposon screens, including Spic, which encodes a PU.1-related transcription factor, and Hdac7, a histone deacetylase gene. PiggyBac and Sleeping Beauty have different integration preferences. To maximize the utility of the tool, we engineered 20 mouse lines to be compatible with both transposases in constitutive, tissue-, or temporal-specific mutagenesis. Mice with ...
The extraintestinal pathogen, avian pathogenic E. coli (APEC), known to cause systemic infections in chickens, is responsible for large economic losses in the poultry industry worldwide. In order to identify genes involved in the early essential stages of pathogenesis, namely adhesion and colonization, Signature-tagged mutagenesis (STM) was applied to a previously established lung colonization model of infection by generating and screening a total of 1,800 mutants of an APEC strain IMT5155 (O2:K1:H5; Sequence type complex 95). The study led to the identification of new genes of interest, including two adhesins, one of which coded for a novel APEC fimbrial adhesin (Yqi) not described for its role in APEC pathogenesis to date. Its gene product has been temporarily designated ExPEC Adhesin I (EA/I) until the adhesin-specific receptor is identified. Deletion of the ExPEC adhesin I gene resulted in reduced colonization ability by APEC strain IMT5155 both in vitro and in vivo. Furthermore, complementation of
In this study, we created a fission yeast insertion mutant library in which all mutants were tagged with unique barcode sequences and stored as two readily available selection platforms. The 384-well mutant arrays allow genetic screens on individual mutants and can be extended to genetic approaches such as synthetic genetic array (SGA) [45, 46]. These mutant arrays have been used to identify mutants with four distinct phenotypes (Table 2) as well as strains that are hyper-sensitive to cancer chemotherapeutics camptothecin and bleomycin (Hale and Runge, unpublished data). In addition to 384-well mutant arrays, mutant pools of 1800 mutants are available for parallel analysis.. The insertion mutagenesis used in this study relied on random non-homologous recombination, where a vast majority of transformants have unstable, circularized vector DNA and only a small portion have stable insertions. To facilitate the collection of stable insertion mutants, we included low-dose 5-FOA in our initial ...
In this communication, we report a novel strategy for the genetic manipulation of large viral DNA genomes. In a single step we cloned an infectious cytomegalovirus DNA as a bacterial artificial chromosome in E. coli and reconstituted virus progeny after transfection of the BAC plasmid into eukaryotic cells. This approach makes the CMV genome accessible to the genetic techniques established for E. coli. As an example for the power of the mutagenesis procedures, we performed a targeted insertion of four nucleotides into the 230-kb MCMV genome. In principle, any mutation (point mutations, insertions, and deletions) in any region of the genome can now be introduced using the described mutagenesis procedure. Moreover, other procedures, for example a random transposon mutagenesis of the CMV genome are conceivable. Multiple mutations can be introduced in consecutive rounds of mutagenesis without the need to reconstitute infectious viral intermediates. Construction of revertant genomes can be easily ...
Active retrotransposons play important roles during evolution and continue to shape our genomes today, especially in genetic polymorphisms underlying a diverse set of diseases. However, studies of human retrotransposon insertion polymorphisms (RIPs) based on whole-genome deep sequencing at the population level have not been sufficiently undertaken, despite the obvious need for a thorough characterization of RIPs in the general population.|br| Herein, we present a novel and efficient computational tool named Specific Insertions Detector (SID) for the detection of non-reference RIPs. We demonstrate that SID is suitable for high depth whole-genome sequencing (WGS) data using paired-end reads obtained from simulated and real datasets. We construct a comprehensive RIP database using a large population of 90 Han Chinese individuals with a mean 68× depth per individual. In total, we identify 9342 recent RIPs, and 8433 of these RIPs are novel compared with dbRIP, including 5826 Alu, 2169 long interspersed
A fixed gene copy number is important for the in silico construction of engineered synthetic networks. However, the copy number of integrated genes depends on their genomic location. This gene dosage effect is rarely addressed in synthetic biology. Two studies in Escherichia colipresented conflicting data on the impact of gene dosage. Here, we investigate how genome location and gene orientation influences expression in Bacillus subtilis. An important difference with the E. coli studies is that we used an unbiased genome integration approach mediated by random transposon insertion. We found that there is a strong gene dosage effect in fast growing B. subtilis cells, which can amount to a 5-fold difference in gene expression. In contrast, gene orientation with respect to DNA replication direction does not influence gene expression. Our study shows that gene dosage should be taken into account when designing synthetic circuits in B. subtilis and presumably other bacteria.. ...
Institutions: Samuel Lunenfeld Research Institute We are performing gene trap-based expression and genotypic screens to generate new mouse mutations that will help delineate the molecular controls of specific developmental programs. Using a polyA trap vector with recombination sites for post-insertional manipulations, gene trap insertions are screened using multiplexed in vitro differentiation and induction assays. An expression profile is being generated at a rate of greater than 5000 per year. Sequence tags for all insertions demonstrating restricted expression patterns (about 20%) are now being generated. A database is being developed that will be searchable by expression pattern, sequence, and phenotype. The clones will be available as a resource to researchers worldwide. One of the clones identified in our ongoing screen, SNAG-1, is expressed by hematopoietic stem cells, neural endothelial cells, cardiomyocytes and sensory nerves. SNAG-1 mutant embryos die mid-gestation with neural vascular ...
The mini-Tn5 are mobile elements able to transpose to the chromosome from a delivery plasmid, by means of the transposase activity encoded by the tnp* gene, which is present in cis in the plasmid but external to the mini-Tn5 element. pUT mini-Tn5 vectors can be used to insert any DNA fragment in the bacterial chromosome, by cloning it within the mini-Tn5 region. Due to the loss of the tnp* gene after insertion, minitransposons are stably integrated into the chromosome and inherited. The different pUTmini-Tn5 vectors can then be used for repeated insertion events and therefore multiple insertions in the same strain are only limited by the availability of distinct selection markers.. Advantages:. ...
Atarés Huerta, Alejandro; Moyano, Elena; Morales, Belén; Schleicher, Peter; García Abellán, José Osvaldo; Antón Martínez, María Teresa; García Sogo, Begoña; Pérez Martin, Fernando; Lozano, Rafael; Borja Flores, Francisco; Moreno Ferrero, Vicente; Bolarin Jimenez, Maria del Carmen; Pineda Chaza, Benito José (Springer Verlag (Germany), 2011) ...
A new paper in Cell Reports utilizes RNA-seq and Tn-seq (the "tn" in tn-seq stands for transposon) to map the transcriptional and fitness changes in bacterial gene networks in response to stressors, like nutrient depletion and antibiotics.. The transcriptional response measures changes in gene expression as measured by RNA-seq. The fitness or phenotypic response describes the importance of each gene to the response. This is measured by a different assay, Tn-seq, which takes advantage of transposon insertion to selectively inactivate genes in the bacterial genome. Those genes that are depleted in the stressor condition are determined to be "high fitness" (owing to the fact that the bacteria without those genes died under stress).. First, before even considering DE genes, they found that there is no correlation between a genes transcriptional abundance (not fold change) and its fitness. While most high-fitness genes were also high abundance, many more high-abundance genes were not high-fitness. ...
A DNA transposon, or jumping gene, controls its amplification within a genome through a competition between the enzyme multimers that are responsible for its mobility.
Interestingly there is no rule such as the higher evolved an organism the more transposable elements. Although we find more transposable elements in higher evolved organisms this rule can not be maintained if compared single species as frogs and humans.. Transposons most often code for transposase the enzyme responsible for the transposon dislocation. As Transposons are so common in a genome it is not surprising that transposases are the most common genes in a genome [2].. ...
Hard hitting drums and a huge bass lead that will have you up in arms and ready to defend New York from the Cleaners and LMB! Equip those new mods and party up! WERE HEADED TO THE DARK ZONE!. ...
The Rosa26 gene locus is well suited for gene over-expression. Reduced development time and cost with ready-to-use targeting vector.
Dr. performed reduction and percutaneous rush pins insertion on open fracture of humeral neck. What is a rush pin considered? What code wo
A summary of The Insertion Sort Algorithm in s Insertion Sort. Learn exactly what happened in this chapter, scene, or section of Insertion Sort and what it means. Perfect for acing essays, tests, and quizzes, as well as for writing lesson plans.
It is possible that there is an internal rearrangement of the MiMIC transposon involving the 3prime end. May be segregating SM6a ...
Saha, N.,Tay, J.S.H.,Basair, J.,Talmud, P.J.,Humphries, S.E. (1996). Lack of association of angiotensin-converting enzyme (ACE). Gene insertion/deletion polymorphism with CAD in two Asian populations. Clinical Genetics 50 (3) : 121-125. [email protected] Repository ...
In this study, first we showed that the Tol2 transposon system is a useful technique in generating stable transfected primary culture cells and, second, we demonstrated that the Tol2 transposon system is applicable to the study of circadian clock oscillations.. The Tol2 transposon was originally discovered from Medaka fish (Orzyias latipes) [12]. An active autonomous member of Tol2 was first identified by the analysis using zebrafish embryos [13]. Since then, the Tol2 transposon system has been mainly used for random insertion mutagenesis and transgene in zebrafish [14]. Although recent reports have indicated that the transposon systems such as piggyBac and SleepingBeauty in addition to Tol2 are also active in mammalian cells [15, 16], few studies have been reported that utilized the Tol2 system for transfection to mammalian primary culture cells. In the present study, we showed that the Tol2 transposon system is a useful tool in generating stable transfected primary culture cells such as MEF ...
Assigning functions to all 26,000 Arabidopsis genes is a monumental task that will require complementary research strategies and extensive multinational coordination. Part of this effort must be devoted to characterizing gene expression patterns and validating biochemical functions of protein products within the plant cell. A second critical component will be to determine the biological significance of each gene product and its relevance to plant growth and development. This will require forward genetics and saturation mutagenesis on a genomics level. When gene functions are not redundant, this objective can be met by determining which gene disruptions result in a mutant phenotype.. We describe here an efficient system for identifying large numbers of genes with essential and nonredundant functions during seed development. More than 120,000 T-DNA insertion lines were screened for defects in seed development, 4200 seed mutants with a wide range of defects were identified, 1705 confirmed mutants ...
ISHS I International Symposium on Genetic Modifications - Challenges and Opportunities for Horticulture in the World INSERTIONAL MUTAGENESIS IN THE DIPLOID STRAWBERRY (FRAGARIA VESCA)
Here we characterized the sae operon, an independent global regulator for virulence gene expression in S. aureus. Sequence analysis revealed two additional ORFs upstream of the two-component system saeRS. Both ORFs are predicted to code for putative proteins with yet unknown functions. The predicted protein encoded by ORF3 is probably membrane associated, and that encoded by ORF4 is probably cytosolic. Transciptional analysis leads to the assumption that both ORFs are functionally linked to the saeRS two-component regulatory system. ORF3 is cotranscribed with saeRS in the major transcripts T1 and T2. ORF4 is cotranscribed with ORF3 and saeRS (T1) but is also contained in the monocistronic T4 transcript. All transcripts, including the monocistronic message T4, are absent in the sae mutant strain, although the transposon insertion site was shown to be localized further downstream within saeS. Thus, SaeRS is probably necessary for transcriptional initiation from P1. Transcriptional initiation from ...
In this investigation, we have used transposon mutagenesis to identify a novel mycobacterial protein that is associated with phenotypic changes in the parent strain that alter the adhesive properties of the mycobacteria. Recently, a similar BCG transposon library was used to identify a mycolic acid cyclopropane synthetase as a virulence factor in M. tuberculosis by screening for variants that are deficient in cord formation (20). For our investigation, we reasoned that a mutant which exhibited a nonaggregative morphology may identify a gene that influences the interaction of mycobacteria with other cells, since there is evidence that adhesins can also induce autoaggregation of bacteria (9, 32). In fact, it has previously been shown that the mycobacterial adhesin HBHA promotes bacterial aggregation (16, 31, 33).. In this study, after screening more than 1,900 BCG transposon insertion mutants, one mutant strain, mc21525, was singled out for demonstrating an obvious difference in clumping ...
SB transposon mutagenesis is an unbiased approach for identifying candidate BC driver genes. We successfully induced mammary tumors in mice using the K5 promoter driving SB alone or together with stabilized N-terminally truncated β-catenin targeted to the basal layer of the mammary gland (28). Because the K5-Cre promoter is activated in both the luminal and basal cell layers of the mammary gland, transposition also occurs in both layers and not solely in the basal cell layer, as initially observed with the transgenic line expressing the truncated β-catenin. Not surprisingly, mammary tumors induced by our SB system represented all BC histological subtypes, consistent with the premise that the cell of origin for BC derives from either the luminal or basal layers of mammary glands (56, 57).. The SB mouse model provides a unique experimental basis for the identification of BC-associated susceptible genes relevant to the tumor subtypes. To understand the molecular subtypes of tumors induced in our ...
Drug resistance poses a great challenge to targeted cancer therapies. In Hedgehog pathway-dependent cancers, the scope of mechanisms enabling resistance to SMO inhibitors is not known. Here, we performed a transposon mutagenesis screen in medulloblastoma and identified multiple modes of resistance. Surprisingly, mutations in ciliogenesis genes represent a frequent cause of resistance, and patient datasets indicate that cilia loss constitutes a clinically relevant category of resistance. Conventionally, primary cilia are thought to enable oncogenic Hedgehog sig-naling. Paradoxically, we find that cilia loss protects tumor cells from susceptibility to SMO inhibitors and maintains a "persister" state that depends on continuous low output of the Hedgehog program. Persister cells can serve as a reservoir for further tumor evolution, as additional alterations synergize with cilia loss to generate aggressive recurrent tumors. Together, our findings reveal patterns of resistance and provide mechanistic ...
Transposable elements (TEs) are an important source of genomic variability in eukaryotic genomes. Their activity impacts genome architecture and gene expression and can lead to drastic phenotypic changes. Therefore, identifying TE polymorphisms is key to better understand the link between genotype and phenotype. However, most genotype-to-phenotype analyses have concentrated on single nucleotide polymorphisms as they are easier to reliable detect using short-read data. Many bioinformatic tools have been developed to identify transposon insertions from resequencing data using short reads. Nevertheless, the performance of most of these tools has been tested using simulated insertions, which do not accurately reproduce the complexity of natural insertions. We have overcome this limitation by building a dataset of insertions from the comparison of two high-quality rice genomes, followed by extensive manual curation. This dataset contains validated insertions of two very different types of TEs, LTR
A Genetic Screen Using the PiggyBac Transposon in Haploid Cells Identifies Parp1 as a Mediator of Olaparib Toxicity. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
Advenviral Vector - Gene Insertion Nucleus Tumor Cell , Gene insertion nucleus tumor cell. Adenovirus inserting gene into tumor cell nucleus. Nuclear por, nucleus, episome, DNA, mRNA, ribosome, protein, transcription, translation, oncology, immunity .
The transposable element (TE), Tn5, is a conservative transposon that is able to insert a segment of genes bordered by specific 19bp insertion sequences (IS) from one part of the genome (e.g. plasmid vector) randomly to another location, such as the chromosome (Reznikoff, 2008). The transposition event is catalyzed by a transposase enzyme encoded by Tnp gene included in the TE. By inserting a vector construct containing the TE with selectable markers (such as tetracyclin resistance and lacZ) into an organism with a desirable phenotype, we can find out what genetic elements (e.g. genes and promoters) are responsible for that particular function. This can happen via a random insertion of a TE containing a promoterless reporter gene downstream of promoter elements that creates a transcriptional fusion, providing activity in response to specific environmental stimuli. Another advantage of using a transposon approach is that it creates a saturating library of mutants where all possible genetic ...
Shoko was a postdoctoral fellow at the Nara Institute, In Japan, in 2001 and joined the Amaya lab in 2002. Originally based at the Wellcome Trust/Cancer Research UK Gurdon Institute in Cambrdige, Shoko relocated to the University of Manchester with the Amaya lab in 2006. Shoko is currently working on the development of the gene trap transposon system for insertional mutagenesis in Xenopus tropicalis ...
pUT mini-Tn5 System. The mini-Tn5 are mobile elements able to transpose to the chromosome from a delivery plasmid, by means of the transposase activity encoded by the tnp* gene, which is present in cis in the plasmid but external to the mini-Tn5 element. pUT mini-Tn5 vectors can be used to insert any DNA fragment in the bacterial chromosome, by cloning it within the mini-Tn5 region. Due to the loss of the tnp* gene after insertion, minitransposons are stably integrated into the chromosome and inherited. The different pUTmini-Tn5 vectors can then be used for repeated insertion events and therefore multiple insertions in the same strain are only limited by the availability of distinct selection markers.. Advantages:. ...
The KOMP Repository Collection is located at the MMRRC at the University of California, Davis and Childrens Hospital Oakland Research Institute. Question? Comments? For Mice, Cells, and germplasm please contact us at [email protected], US 1-888-KOMP-MICE or International +1-530-752-KOMP, or for vectors [email protected] or +1-510-450-7917 ...
This classic human skeleton model (we call him Stan) has been the standard anatomical skeleton model of quality in hospitals, schools, universities, and...
Such Knockin mice have an inserted exogenous gene that is regulated under the native promoter, providing a physiologically relevant and secure expression.
In the past five years we have identified the developmental stage of Tcell differentiation at which malignant transformation caused by retroviral insertion of a...
Colletotrichum lindemuthianum is one of the main pathogens affecting common bean (Phaseolus vulgaris) in tropical regions. In the present work we describe three mutants of C. lindemuthianum obtained by insertional mutagenesis using the REMI technique (Restriction Enzyme-Mediated Integration). The mutants were selected based on their lower virulence/pathogenicity and the partial characterization of the mutated genes is presented. In addition, the mitochondrial genome of this phytopathogen was partially characterized. The results show that the gene pacC1 plays a role in vegetative growth and pathogenicity. The use of the restriction enzyme NarI in the transformation of C. lindemuthianum increased approximately 10-fold the transformation efficiency using plasmid pAN7.1. Mutant strains mut5, mut29, and mut65 were isolated from a total of 580 transformants based on their altered ability to infect susceptible bean plants. Strains mut29 and mut65 failed to penetrate hypocotyl tissues of the common ...
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Plasmodium falciparum is a human intracellular parasite that is the causative agent of a deadly form of malaria. This species alone is responsible for 200 million cases of malaria annually resulting in over 1 million deaths worldwide. The excessive mortality due to P. falciparum infection is due to its ability to cause severe pathogenesis through hyperparasitemia and cytoadherence defined as the ability of infected red blood cells to adhere to host vasculature. Cytoadherence is mediated through the export of parasite proteins to the surface of the infected red blood cell (RBC). Exported proteins have been identified but the pathway for protein export is still being elucidated. Many protein coding genes in the P. falciparum genome are hypothetical and therefore still need to be studied. Random transposon mutagenesis using the piggyBac transposable element in P. falciparum has given us a library of mutants to use for forward genetic studies. In this work, we describe a novel approach for screening P.
In any transgenic mice, insertion of the transgene in the genome is potentially mutagenic. It has been proposed that about 5-10% of transgenic mice harbour transgene induced chromosomal alterations, including deletions and translocations.7,8 Thus, random insertion of foreign DNA into any genome is liable to deactivate or activate one or more genes. The striking eye phenotype observed in the T27aT15 transgenic line seems to be the result of the transgene insertion into the genome as the phenotype is unique among more than twenty transgenic lines that were generated using identical or similar constructs.3. We present a phenotypic characterisation of the T27aT15 line. A proportion of hemizygous transgenic mice developed serious abnormalities of the eye, usually starting between one and two months of age. The progressive eye abnormalities seem to arise after eye formation, as the eye appears to develop normally. The phenotype of these mice includes corneal opacity, cataract, and retinal vascular ...
b)The manuscripts listed below should be referred in the publication on scientific journal.(1)Ito T, Motohashi R, Kuromori T, Mizukado S, Sakurai T, Kanahara H, Seki M and Shinozaki K (2002) A new resource of locally transposed Dissociation elements for screening gene-knockout lines in silico on the Arabidopsis genome. Plant Physiol. 129: 1695-1699.(2)Kuromori T, Hirayama T, Kiyosue Y, Takabe H, Mizukado S, Sakurai T, Akiyama K, Kamiya A, Ito T and Shinozaki K (2004) A collection of 11800 single-copy Ds transposon insertion lines in Arabidopsis. Plant J. 37, 897-905 ...
Despite the wide range of techniques that can be brought to bear on the study of basic processes in Drosophila, there are still deficiencies in our armory. One of these is an ability to select mutants in cases where the gene is known and has been cloned, but where we are ignorant of the associated phenotype. We describe here a solution to this problem as applied to a model system, the singed (sn) locus. Our method is a combination of classical genetics and molecular biology: sib selection plus the polymerase chain reaction. We have used the method to isolate rare individuals with P-element-induced alleles of sn merely by recognition of the DNA structures induced at the locus by transposon insertion. Phenotypic criteria were used only retrospectively to verify our diagnoses. There are obvious implications of this technique for the mutagenesis of other organisms.
Characterization of a gene trap insertion into a novel gene, cordon-bleu, expressed in axial structures of the gastrulating mouse embryo.. We have used a gene trap (GT) vector and embryonic stem (ES) cell chimeras to screen for insertions of the lacZ reporter gene into transcription units that are spatially and temporally regulated during early mouse embryogenesis. GT vectors which can act as both a reporter and a mutagen have been previously used to isolate new genes that are essential for mouse development. In this paper we describe a GT insertion which displays a very restricted pattern of expression in the gastrulating embryo. beta-Galactosidase activity was first detected at 7.5 days post-coitum (E7.5) in the node region of the embryo and extended to the midline structures at E8.0. At E9.5 expression was restricted to the floor plate, the notochord, the roof of the gut, and the liver anlage. Expression appeared in the somites at E10.0 and later became more widespread. We used rapid ...
We screened the nonredundant PA14 transposon mutant library for mutants defective in growth on GB to identify genes involved in the GB catabolic pathway in P. aeruginosa. We identified a putative GB demethylase, encoded by gbcA and gbcB, based on phenotypic data that indicated that the ΔgbcA-gbcB mutant could not grow on GB but could use DMG as a carbon and nitrogen source (Table 2 and data not shown). Furthermore, when the the ΔgbcA-gbcB mutant was fed choline, GB accumulated in the cells (Fig. 4). The gbcA and gbcB transcript levels increased in response to GB and DMG in a GbdR-dependent manner. The transcript accumulation was mirrored in a proteomics analysis, in which the GbcB protein was shown to be more abundant in P. aeruginosa grown in the presence of GB as the sole carbon source (11). We also identified a putative DMG demethylase, encoded by the dgcAB genes, which is necessary for conversion of DMG to sarcosine. Experiments with 13C-labeled choline confirmed that DMG accumulated in ...
PiggyBac Transposable Element Derived 5 is an enzyme that in humans is encoded by the PGBD5 gene.[1] PGBD5 is a DNA transposase related to the ancient PiggyBac transposase first identified in the cabbage looper moth, Trichoplusia ni.[2] The gene is believed to have been domesticated over 500 million years ago in the common ancestor of cephalochordates and vertebrates.[3] The putative catalytic triad of the protein composed of three aspartic acid residues is conserved among PGBD5-like genes through evolution,[4], and is distinct from other PiggyBac-like genes.[3] PGBD5 has been shown to be able to transpose DNA in a sequence-specific, cut-and-paste fashion.[4] PGBD5 has also been proposed to mediate site-specific DNA rearrangements in human tumors.[5] ...
Cell culture and transposition assay HEK 293 cells had been maintained in MEMa medium supplemented with 10% FBS, 100 units ml penicillin, and a hundred ug mL streptomycin. The specifics for your transposition assays had been described pre viously. Inhibitors,Modulators,Libraries Exercise assay in the piggyBac transposase A equivalent procedure as thorough previously was employed to co transfect a hundred ng of piggyBac donor, with various quantity of the piggyBac helper, pCMV Myc piggyBac, ranging from 0 300 ng into 1. 2 105 of HEK 293 cells. pcNDA3. 1NEO, an empty vector used in our prior examine, was employed to best the total level of DNA transfected to 400 ng. Each and every trans fection condition was completed in triplicate. Twenty 4 hours just after transfection, one fifth of transfected cells were subjected to transposition assay.. The remaining transfected cells in triplicate had been pooled and grew in the 35 mm plate for a further twenty four hours prior to staying subjected to ...
Joel Neal MD Of Stanford University Medical Center Discusses What Is EGFR Exon 20 Insertion Mutant NSCLC: 15% Of Lung Cancer In The United States, 50% Overall In Asian Countries. - Non-Small Cell Lung Cancer
Advenviral Vector - Gene Insertion Nucleus Tumor Cell,Medical Illustration database of the best portfolios and stock images now features General and Commercial Illustration and illustrators. 8,000+ image database includes all types of subjects and features the largest directory of medical, science, and nature illustrators and illustration on the web.
Naturally occurring mutations involving the nervous system have provided virtually all of our current understanding of the genetic regulation of neural development (Caviness and Rakic, 1978). The difficulty of isolating the corresponding genes, however, has precluded a molecular analysis of these mutants. Insertional mutagenesis, induced by microinjection of DNA into fertilized ova to produce transgenic animals, provides a molecular tag that marks the site of the mutational event. In this article, we describe a transgenic neurological mutation, designated wocko (Wo), which disrupts the development of the inner ear. These mutant mice display a dominant behavioral phenotype that consists of circling, hyperactivity, and head tossing, reminiscent of the shaker/waltzer class of mutants, and they display a recessive homozygous sublethal phenotype. Anatomical analyses showed that both structural and neural components of the vestibular system were disrupted, while analyses of mutant fetuses showed that ...
1) a small deletion occurs in the transposase gene of an IS element and plasmid is integrated , 2) a small deletion occurs in the transposase gene of an IS element , 3) two IS elements integrate into a chromosome with only a small distance separating them , 4) an IS element integrates with another IS element with the help of a plasmid
LEAD: Scientists have cured cystic fibrosis cells in the laboratory by inserting a healthy version of the gene that causes the disease, an unexpectedly swift advance that left researchers almost giddy with delight.
Definition of insertion in the Legal Dictionary - by Free online English dictionary and encyclopedia. What is insertion? Meaning of insertion as a legal term. What does insertion mean in law?
LoFric Origo. LoFric®OrigoTM is the instantly-activated male coudé catheter that makes life easier for experienced users and beginners alike. Discreetly packaged and foldable, LoFric Origo is easy to carry and use anywhere you happen to be. Its unique Insertion Grip provides extra control with no need to touch the catheter tube, ensuring safe and hygienic catheterization.. ...
Recombination Mediated Cassette Exchange of a Mi{MIC} insertion results in expression of Egfr tagged with EGFP-FlAsH-StrepII-TEV-3xFlag. May be segregating CyO ...
An insertion system includes a handle assembly and a nose assembly removably attached to the handle assembly and including an insertion end. The handle assembly includes a main body, a nose interface
Learn more about Pacemaker Insertion at St. Davids HealthCare DefinitionReasons for ProcedurePossible ComplicationsWhat to ExpectCall Your Doctorrevision ...
Biomigas site-saturation mutagenesis technology systematically substitutes wildtype amino acids with partial or all 19 non-wildtype mutants.. Advantages. ...
Sometimes in the middle of a heated, political squabble that has drug on for many months, you can feel the need to just get into the nearest cave and wait until it all blows over. Compounding that is the fact that as I write this its the month of February...
A versatile genetic method for identifying and cloning Drosophila melanogaster genes affecting any recognizable phenotype is described. Strains are constructed in which the insertion of a single P transposable element has caused a new mutation, greatly simplifying the genetic and molecular analysis of the affected gene. Mutagenesis is initiated by crossing two strains, each of which contains a specially designed P element. One element (jumpstarter), encoding P element transposase, efficiently mobilizes the second nonautonomous transposon (mutator), whose structure facilitates selection and cloning of new insertion mutations. Random mutator transpositions are captured in individual stocks that no longer contain jumpstarter, where they remain stable. This method was used to construct 1300 single P element insertion stocks which were then screened for recessive mutations. A library of single-element insertion strains will allow the structure and function of Drosophila genes to be readily ...
Malignant primary brain tumors, gliomas, often overexpress both platelet-derived growth factor (PDGF) ligands and receptors providing an autocrine and/or paracrine boost to tumor growth. Glioblastoma multiforme (GBM) is the most frequent glioma. Its aggressive and infiltrative growth renders it extremely difficult to treat. Median survival after diagnosis is currently only 12-14 months. The present review describes the use of retroviral tagging to identify candidate cancer-causing genes that cooperate with PDGF in brain tumor formation. Newborn mice injected intracerebrally with a Moloney murine leukemia retrovirus carrying the sis/PDGF-B oncogene and a replication competent helper virus developed brain tumors with many characteristics of human gliomas. Analysis of proviral integrations in the brain tumors identified almost 70 common insertion sites (CISs). These CISs were named brain tumor loci and harbored known but also putative novel cancer-causing genes. Microarray analysis identified ...
Legionella pneumophila is a facultative intracellular bacillus that causes nosocomial and community-acquired pneumonia and, rarely, extrapulmonary infections in humans. Signature-tagged mutagenesis employs uniquely tagged transposons that are used to randomly mutagenize a bacterial chromosome and create a library. A library of 700 mutant clones created by signature-tagged mutagenesis was screened using this negative-selection strategy in Acanthamoeba castellanii, a free-living amoeba that may serve as an environmental reservoir of legionellae. The efficiency of invasion was studied by incubating L. pneumophila strains grown to postexponential phase with A. castellanii, using gentamicin to kill extracellular organisms and then determining remaining intracellular CFU. The behavior of the mutants was also examined in human macrophages derived from peripheral blood mononuclear cells (PBMCs) and in phorbol myristate acetate (PMA)-differentiated U-937 cells. In contrast, all of the mutants replicated within
Several studies have suggested that Insertion/Deletion polymorphism of ApoB gene is associated with obesity, dyslipidemia, diabetes and coronary heart disease (CHD).
A number of genetic determinants required for bacterial colonization of solid surfaces and biofilm formation have been identified in different micro-organisms. There are fewer accounts of mutations that favour the transition to a sessile mode of life. Here we report the isolation of random transposon Pseudomonas putida KT2440 mutants showing increased biofilm formation, and the detailed characterization of one of them. This mutant exhibits a complex phenotype, including altered colony morphology, increased production of extracellular polymeric substances and enhanced swarming motility, along with the formation of denser and more complex biofilms than the parental strain. Sequence analysis revealed that the pleiotropic phenotype exhibited by the mutant resulted from the accumulation of two mutations: a transposon insertion, which disrupted a predicted outer membrane lipoprotein, and a point mutation in lapG, a gene involved in the turnover of the large adhesin LapA. The contribution of each ...
Genome-wide gene insertion and deletion rates can be modelled in a maximum likelihood framework with the additional flexibility of modelling potential missing data using the models included within. These models simultaneously estimate insertion and deletion (indel) rates of gene families and proportions of missing data for (multiple) taxa of interest. The likelihood framework is utilized for parameter estimation. A phylogenetic tree of the taxa and gene presence/absence patterns (with data ordered by the tips of the tree) are required. For more details, see Utkarsh J. Dang, Alison M. Devault, Tatum D. Mortimer, Caitlin S. Pepperell, Hendrik N. Poinar, G. Brian Golding (2016). Gene insertion deletion analysis while accounting for possible missing data. Genetics (accepted).
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We have developed a new strategy designated SIMF (Systematic Insertional Mutagenesis of Families), to identify DNA insertions in many members of a gene family simultaneously. This method requires only a short amino acid sequence conserved in all members of the family to make a degenerate oligonucleotide, and a sequence from the end of the DNA insertion. The SIMF strategy was successfully applied to the large maize R2R3 Myb family of regulatory genes, and Mutator insertions in several novel Myb genes were identified. Application of this technique to identify insertions in other large gene families could significantly decrease the effort involved in screening at the same time for insertions in all members of groups of genes that share a limited sequence identity.. ...
Name: SALK_063013. ABRC stock number: SALK_063013. Description: Sequence-indexed T-DNA insertion line generated by vacuum infiltration of Columbia (Col) plants with Agrobacterium tumefaciens vector pROK2; kanamycin was employed for selection of plants carrying a T-DNA (please note that in many lines, the kan resistance trait has been cosuppressed, so that insertion plants may not be kanamycin resistant); each T1 transformant has been maintained individually at SALK; the DNA sequence of each T-DNA flanking region was generated from seedlings grown from the same sample of seeds as that provided for distribution (T3). NOTE: kanamycin resistance gene may be silenced; PCR- or hybridization-based segregation analysis is required to confirm presence of insertion; may be segregating for phenotypes that are not linked to the insertion; may have additional insertions potentially segregating. Please cite the Alonso et al. reference linked to this stock and acknowledge NASC/ABRC for distributing the seeds ...
Rice, Oryza sativa L., is one of the most important crops in the world. With the rising world population, feeding people in a more sustainable and environment-friendly way becomes increasingly important. Therefore, rice research community needs to share resources to better understand functions of rice genes that are the foundation for future agricultural biotechnology development, and one way to achieve this goal is via the extensive study of insertional mutants.|br| We have constructed a large rice insertional mutant population in a japonica rice variety, Tainung 67. The collection contains about 93,000 mutant lines, among them 85% with phenomics data and 65% with flanking sequence data. We screened the phenotypes of 12 individual plants for each line grown under field conditions according to 68 subcategories and 3 quantitative traits. Both phenotypes and integration sites are searchable in the database at Taiwan Rice Insertional Mutants Database (http://trim.sinica.edu.tw).|br| Detailed analyses of
Rice, Oryza sativa L., is one of the most important crops in the world. With the rising world population, feeding people in a more sustainable and environment-friendly way becomes increasingly important. Therefore, rice research community needs to share resources to better understand functions of rice genes that are the foundation for future agricultural biotechnology development, and one way to achieve this goal is via the extensive study of insertional mutants.|br| We have constructed a large rice insertional mutant population in a japonica rice variety, Tainung 67. The collection contains about 93,000 mutant lines, among them 85% with phenomics data and 65% with flanking sequence data. We screened the phenotypes of 12 individual plants for each line grown under field conditions according to 68 subcategories and 3 quantitative traits. Both phenotypes and integration sites are searchable in the database at Taiwan Rice Insertional Mutants Database (http://trim.sinica.edu.tw).|br| Detailed analyses of
misc{7861784, abstract = {DNA transposons are a class of mobile genetic elements that can autonomously move from one genomic location to another. They are powerful drivers of genetic change and have played a significant role in the evolution of many genomes. One such transposable element, IS608 from Helicobacter pylori employs a unique mechanism of transposition as it transposes in a single-stranded DNA form and inserts specifically 3 of a specific tetranucleotide sequence (Kersulyte et al., 2002; Guynet et al., 2008). Previous structural (Ronning et al., 2005; Barabas et al, 2008) and biochemical studies (Ton-Hoang et al., 2005) of IS608, revealed that the element chooses its integration site specifically via base-pairing between the transposon and the target DNA. This unique feature allowed re-directing transposon integration to various four-nucleotide sequences by simply modifying the transposon DNA sequence. A key feature of the retargeting strategy was that, unlike the previous attempts to ...
Despite the recent large-scale efforts dedicated to comprehensive phylogenetic analyses using mitochondrial and nuclear DNA sequences, several relationships among mammalian orders remain controversial. Here, we present an extensive application of retroposon (L1) insertion analysis to the phylogeneti …
Until recently, a transposon popularly known as Sleeping Beauty was thought to be the most efficient in inserting genes into mammalian systems. However, a drawback to Sleeping Beauty is its inability to be modified to overcome concerns about placing therapeutic genes safely within the genome. Moisyadi and Kaminski compared a number of transposons and found that piggyBac is much more efficient at inserting transgenes into the genome of several human lines. It is also amenable to molecular alteration so that it can potentially insert a therapeutic gene into a safe area of the genome. In fact, the piggyBac transposase enzyme that performs the insertion of the transposon into DNA can now be modified to attempt placing the transgenes into pre-designated regions of the genome - targeted insertion ...