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  • Zinc Finger Nu
  • The frequency of gene targeting can be significantly enhanced through the use of engineered endonucleases such as zinc finger nucleases, engineered homing endonucleases, and nucleases based on engineered TAL effectors. (wikipedia.org)
  • To date, four classes of SSN -meganucleases (homing endonuclease), zinc finger nucleases ( ZFN s), transcriptional activator-like effector nucleases ( TALENs ), and the clustered regularly interspaced short palindromic repeat ( CRISPR )/ CRISPR -associated protein 9 ( Cas9 )-have been developed to cleave genes of interest. (plantphysiol.org)
  • Zinc finger nucleases have also been used in a mouse model of haemophilia and a clinical trial found CD4+ human T-cells with the CCR5 gene disrupted by zinc finger nucleases to be safe as a potential treatment for HIV/AIDS. (wikipedia.org)
  • genome
  • Following infection with Agrobacterium , transfer of transfer DNA ( T-DNA ) to the nucleus and its integration into the plant genome occur consecutively during cocultivation, thus timing the induction of DNA double-strand breaks ( DSB s) on the target gene to coincide with the delivery of the HDR template is crucial. (plantphysiol.org)
  • Genetic modification, such as the addition of exogenous genes to the MSC genome, is crucial to their use as cellular vehicles. (hindawi.com)
  • Neo gene from Tn5, which confers resistance to kanamycin in bacteria and geneticin in eukaryotic cells Mutant FabI gene (mFabI) from E. coli genome, which confers triclosan resistance to the host. (wikipedia.org)
  • The targeted deletion or alteration of moss genes relies on the integration of a short DNA strand at a defined position in the genome of the host cell. (wikipedia.org)
  • Genome browser BLAST Expression Search and Clone Search (search by gene symbol, gene name, or Affymetrix id) Gene nomenclature guidelines Literature search: Textpresso- uses an algorithm to match your search to specific criteria or section of a paper. (wikipedia.org)
  • mutations
  • The method can be used to delete a gene, remove exons, add a gene, and introduce point mutations. (wikipedia.org)
  • Gene targeting has been widely used to study human genetic diseases by removing ("knocking out"), or adding ("knocking in"), specific mutations of interest to a variety of models. (wikipedia.org)
  • The ability to specifically modify the genomes of yeast and mouse provides a quick and easy way to generate or rescue mutations in genes for which a DNA clone or sequence is available. (sciencemag.org)
  • however, not all genes are mutated in such collections, and often the mutations that are generated are not null alleles. (sciencemag.org)
  • TO discover the function of a gene, a geneticist typically isolates mutations in that gene and evaluates their effects. (genetics.org)
  • It is frequently desirable to be able to introduce targeted mutations to address specific questions about gene function. (genetics.org)
  • Particularly with the sequences of complete genomes of many organisms now available, gene identification is facilitated, but producing directed mutations is often still challenging. (genetics.org)
  • The panel provides Watson information on amino acid mutations -- amino acids are a major component and factor in the function of the proteins our genes make and so their normal and cancer-promoting functions. (eurekalert.org)
  • It also provides known gene fusions or mutations, like those with BRAF, which can transform previously inactive genes into oncogenes on eight common cancers, like lung, colon, breast, prostate as well as melanoma. (eurekalert.org)
  • As an example, as one of many test runs, Watson provided Kolhe a 30-page assessment of a patient with a rare soft tissue sarcoma, that included mutations on seven genes, where the mutations are, what they are known to do and options for targeting them. (eurekalert.org)
  • There also was a list of gene alterations that have no known therapies as well as gene mutations that were present, but their contribution to disease aren't currently known. (eurekalert.org)
  • They realised independently that a technique used to mutate bacteria through exposing them to x-rays could also introduce mutations into mammalian genes. (animalresearch.info)
  • Mutations in the COL1A1 gene are associated with the following conditions: Ehlers-Danlos syndrome, arthrochalasia type is caused by mutations in the COL1A1 gene. (wikipedia.org)
  • The mutations in the COL1A1 gene that cause this disorder instruct the cell to leave out a part of the pro-alpha1(I) chain that contains a segment used to attach one molecule to another. (wikipedia.org)
  • Osteogenesis imperfecta, type I: Osteogenesis imperfecta is the most common disorder caused by mutations in this gene. (wikipedia.org)
  • Mutations that inactivate one of the two copies of the COL1A1 gene cause osteogenesis imperfecta type I. The mutated copy of the gene does not produce any pro-alpha1(I) collagen chains. (wikipedia.org)
  • Osteogenesis imperfecta, type II: Many different types of mutations in the COL1A1 gene can cause osteogenesis imperfecta type II. (wikipedia.org)
  • These mutations range from missing pieces of the COL1A1 gene to amino acid substitutions, in which the amino acid glycine is replaced by another amino acid in the protein strand. (wikipedia.org)
  • Osteogenesis imperfecta, type III: Mutations in the COL1A1 gene may result in the production of a protein that is missing segments, making it unusable for collagen production. (wikipedia.org)
  • These mutations may involve missing pieces of the COL1A1 gene or changes in base pairs (the building blocks of DNA). (wikipedia.org)
  • proteins
  • These proteins can bind promoter sequences in the host plant and activate the expression of plant genes that aid bacterial infection. (wikipedia.org)
  • When injected into plants, these proteins can enter the nucleus of the plant cell, bind plant promoter sequences, and activate transcription of plant genes that aid in bacterial infection. (wikipedia.org)
  • Genetic constructs to encode TAL effector-based proteins can be made using either conventional gene synthesis or modular assembly. (wikipedia.org)
  • The adenine-thymine rich hairpin loops of the Borrelia bacteria range in size from 5 kilobase pairs (kb) to over 200 kb and contain the genes responsible for producing a group of major surface proteins, or antigens, on the bacteria that allow it to evade the immune response of its infected host. (wikipedia.org)
  • This makes them a particularly attractive system for testing the ectopic activity of gene products and loss-of-function experiments using antagonizing reagents such as morpholinos, dominant-negatives and neomorphic proteins. (wikipedia.org)
  • Genetics
  • Cold Spring Harbor Laboratory, 1994) and Gene Targeting: A Practical Approach by A.L. Joyner, (Oxford University Press, 1993) both provide an excellent introduction to mouse genetics and techniques covered in the course. (bio.net)
  • This approach is called reverse genetics and it is a powerful and sensitive tool to study the function of genes and, when combined with studies in higher plants such as Arabidopsis thaliana, can be used to study molecular plant evolution. (wikipedia.org)
  • vectors
  • Naturally occurring circular plasmids can be modified to contain multiple resistance genes and several unique restriction sites, making them valuable tools as cloning vectors in biotechnology applications. (wikipedia.org)
  • protein
  • The APOE gene, responsible for encoding a protein called apolipoprotein E, is known to dramatically raise the risk of Alzheimer's. (medicalnewstoday.com)
  • In that very study, Dr. Holtzman uncovered the mechanism by which the APOE protein amplifies Alzheimer's disease-related brain damage in mice, and he suggested that future research should focus on targeting the protein to nip the neurodegenerative process in the bud. (medicalnewstoday.com)
  • A sodium/iodide symporter (NIS), also known as a sodium/iodide cotransporter or solute carrier family 5, member 5 (SLC5A5) is a protein that in humans is encoded by the SLC5A5 gene. (wikipedia.org)
  • The gene chosen was a therapeutic protein to demonstrate the potential of such recombinant DNA technology combined with animal cloning. (wikipedia.org)
  • Plasmid DNA vaccines are genetically engineered to contain a gene which encodes for an antigen or a protein produced by a pathogenic virus, bacterium or other parasite. (wikipedia.org)
  • Collagen, type I, alpha 1, also known as alpha-1 type I collagen, is a protein that in humans is encoded by the COL1A1 gene. (wikipedia.org)
  • This mutation substitutes the amino acid cysteine for the amino acid arginine at position 134 in the protein made by the gene. (wikipedia.org)
  • Sometimes one end of the gene (called the C-terminus) is altered, which interferes with the association of the protein strands. (wikipedia.org)
  • chromosome
  • An alternative arrangement, where DSBs are provided at each end of a homologous segment, is termed ends-out targeting, and causes a segment of chromosome to be replaced with an introduced segment (Fig. 1 ). (pnas.org)
  • The COL1A1 gene is located on the long (q) arm of chromosome 17 between positions 21.3 and 22.1, from base pair 45,616,455 to base pair 45,633,991. (wikipedia.org)
  • sequences
  • Because zinc fingers can be directed to a broad range of DNA sequences and targeting is very efficient, this approach promises to allow genetic manipulation of many different genes, even in cases where the mutant phenotype cannot be predicted. (genetics.org)
  • Generally speaking, expression of a foreign gene requires restructuring the gene to include sequences that are required for producing an mRNA molecule that can be used by the host's translational apparatus (e.g. promoter, translational initiation signal, and transcriptional terminator). (wikipedia.org)
  • Without methods to account for this "context dependence", the standard modular assembly procedure often fails unless it is used to recognize sequences of the form (GNN)N. Numerous selection methods have been used to generate zinc-finger arrays capable of targeting desired sequences. (wikipedia.org)
  • locus
  • Ends-out targeting is more frequently used in mice and yeast because it gives a straightforward route to replace or delete a target locus. (pnas.org)
  • However, when gRNA targeting DNA ligase 4 was transformed with Cas9 prior to the GT experiment, GT efficiency increased dramatically and more than one line exhibiting biallelic GT at the ALS locus was obtained. (plantphysiol.org)
  • The neomycin gene was targeted to the rDNA locus of human MSCs using a nonviral human ribosomal targeting vector. (hindawi.com)
  • Exogenous genes can be targeted to the rDNA locus of human MSCs while maintaining the characteristics of MSCs. (hindawi.com)
  • Both ends of this DNA strand are engineered to be identical to this specific gene locus. (wikipedia.org)
  • plasmid
  • Once delivered into the host, the products of the plasmid genes will then stimulate both the innate immune response and the adaptive immune response of the host. (wikipedia.org)
  • repress
  • Such engineered TAL effectors have been used to create artificial transcription factors that can be used to target and activate or repress endogenous genes in tomato, Arabidopsis thaliana, and human cells. (wikipedia.org)
  • vector
  • Clones were chosen in which the two FRT s were in the same direction, generating the vector pw30, a P element vector carrying a polylinker flanked by I-SceI sites, and then by FRT s, adjacent to a white + ( w + ) gene. (pnas.org)
  • Gene targeting requires the creation of a specific vector for each gene of interest. (wikipedia.org)
  • To synchronize DSB induction and delivery of the HDR template, we transformed a Cas9 expression construct and GT vector harboring the HDR template with guide RNAs ( gRNA s) targeting the rice acetolactate synthase ( ALS ) gene either separately or sequentially into rice calli. (plantphysiol.org)
  • loci
  • Zusammen mit Gene targeting-, die endogene HR Genom eines Organismus an einem bestimmten Loci verändern nutzt, haben HR-basierte Klonierungstechniken erheblich verbessert die Geschwindigkeit und Wirksamkeit von Hochdurchsatz-Gentechnik. (jove.com)
  • Furthermore, this technology enables a robust and efficient Knockin of a particular mutation, reporter, or human gene sequence into endogenous loci, leading to accurate and physiologically more relevant human disease models. (genoway.com)
  • mutation
  • They replaced a specific gene with a similar genetic sequence, which was modified to contain a mutation. (animalresearch.info)
  • Ehlers-Danlos syndrome, classical type: In rare cases, a mutation in the COL1A1 gene has been shown to cause the classical type of Ehlers-Danlos syndrome. (wikipedia.org)
  • sequence
  • This approach, referred to as gene targeting ( GT ), is accomplished by the introduction of DNA fragments encoding a sequence variant of a gene of interest into cells. (plantphysiol.org)
  • The second group deduced the code through molecular analysis of the TAL effector AvrBs3 and its target DNA sequence in the promoter of a pepper gene activated by AvrBs3. (wikipedia.org)
  • expression
  • Increased induction of ZFN expression led to higher frequencies of gene targeting, demonstrating the beneficial effect of activating the target. (genetics.org)
  • These reactions enable four basic functional modules, excision/insertion, inversion, translocation and cassette exchange, which have been used individually or combined in a wide range of configurations to control gene expression. (wikipedia.org)
  • Specific changes to the host organism may be made to improve expression of the ectopic gene. (wikipedia.org)
  • In contrast, growth factors such as IGF-1 and TGF-β (which is induced by the BRAF-V600E oncogene) suppress NIS gene expression, not letting NIS localize in the membrane. (wikipedia.org)
  • resistance
  • Plants have developed a defense mechanism against type III effectors that includes R (resistance) genes triggered by these effectors. (wikipedia.org)
  • targets
  • Gene trapping is based on random insertion of a cassette while gene targeting targets a specific gene. (wikipedia.org)
  • The new targets, rosy ( ry ) and brown ( bw ), both affect eye color and were chosen, like y , for the ease of scoring their mutant phenotypes in these early tests of the generality of ZFN-induced targeting. (genetics.org)
  • reporter
  • Each carried a white + ( w + ) reporter gene and an adjacent I-SceI cut site ( 15 ). (sciencemag.org)
  • One of the tester constructs also carried a partial duplication of the white reporter gene ( Fig. 1 , A and B). As a test for cutting, flies that carried 70I-SceI and a reporter construct were generated by crossing and heat-shocked early in their development. (sciencemag.org)
  • They are a type of reporter gene used in laboratory microbiology, molecular biology, and genetic engineering to indicate the success of a transfection or other procedure meant to introduce foreign DNA into a cell. (wikipedia.org)
  • tissue
  • Additionally, he co-founded Q Therapeutics, a stem-cell research firm and serves on USDA Cellular Tissue and Gene Therapies advisory committee and more. (wikipedia.org)
  • compound
  • The enzyme URA3 also converts 5-fluoroorotic acid (5FOA) into the toxic compound 5-fluorouracil, so any cells carrying the URA3 gene will be killed in the presence of 5FOA (negative selection). (wikipedia.org)
  • Approach
  • In this report we describe our approach more fully, demonstrate its application to two additional genes, and report substantial improvements in targeting efficiency. (genetics.org)
  • copy of the gene
  • This showed how a mouse with a non-functioning copy of the gene differed from normal mice. (animalresearch.info)
  • Because only one copy of the gene is directing the cell to make pro-alpha1(I) chains, cells from people with this disorder make only half of the normal amount of type I collagen, which results in bone fragility and other symptoms. (wikipedia.org)
  • mice
  • A good mouse model for the Fragile X syndrome is available in the form of mice in which the Fmr1 gene (the mouse homologue of the human FMR1 gene) has been invalidated. (wikipedia.org)
  • mutants
  • We demonstrate rapid generation of mutants in seven neuropeptide and two microRNA genes in which no mutants have been described. (genetics.org)
  • The offspring of the host mouse carried the mutant gene, and the offspring of two mutants expressed the mutant gene. (animalresearch.info)
  • yeast
  • In mouse and in yeast some studies show that ends-out targeting is less efficient than ends-in ( 4 - 6 ), whereas others indicate that the two types can be equally efficient ( 7 , 8 ). (pnas.org)
  • pathogenic
  • There also exists virulence plasmids that contain the genetic elements necessary for bacteria to become pathogenic as well as degradative plasmids that harbor the genes that allow bacteria to degrade a variety of substances such as aromatic compounds and xenobiotics. (wikipedia.org)
  • generate
  • Recently, a method to generate such a linear fragment in vivo was reported, accompanied by a demonstration of ends-in or insertional gene targeting ( 3 ). (pnas.org)
  • We constructed a heat-inducible I-SceI gene ( 70I-SceI ) and used standard P element transformation to generate fly lines carrying the transgene ( 14 ). (sciencemag.org)
  • target
  • There are drugs that directly target that mutated gene but if they don't find BRAF, the patient will receive a more broad-spectrum chemotherapy regimen. (eurekalert.org)
  • Watson, on the other hand, has the ability to rapidly identify multiple variations in BRAF along with variations in nearly a dozen other genes known to contribute to the skin cancer as well as the therapies to target them, Kolhe says. (eurekalert.org)
  • Chromatinization of the target DNA inhibits both types of targeting reactions, presumably by impeding RNP access. (utexas.edu)
  • Some of these R genes appear to have evolved to contain TAL-effector binding sites similar to site in the intended target gene. (wikipedia.org)
  • cell
  • Previously used to engineer rat cell models, advances in gene targeting technologies are enabling the creation of a new wave of isogenic human disease models. (wikipedia.org)
  • Complex
  • Examples are the identification of moss genes with implications for crop improvement or human health and the safe production of complex biopharmaceuticals in moss bioreactors. (wikipedia.org)