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MutagenesisMutagenesis, Site-DirectedMutagenesis, InsertionalMolecular Sequence DataAmino Acid SequenceMutationBase SequenceBinding SitesModels, MolecularEscherichia coliAmino Acid SubstitutionStructure-Activity RelationshipSequence Homology, Amino AcidDNA Transposable ElementsRecombinant ProteinsPlasmidsProtein Structure, TertiaryCloning, MolecularBacterial ProteinsKineticsProtein ConformationProtein BindingCysteineProtein Structure, SecondarySequence AlignmentGenes, BacterialPoint MutationMutagensEthylnitrosoureaSubstrate SpecificityCatalysisAlanineDNA PrimersCell LineSOS Response (Genetics)HistidineConserved SequenceDNA Mutational AnalysisCatalytic DomainEscherichia coli ProteinsCrystallography, X-RayGenetic Complementation TestTransfectionRestriction MappingEthyl MethanesulfonateDNA, BacterialSequence DeletionRecombinant Fusion ProteinsProtein EngineeringOligodeoxyribonucleotidesLysinePhenotypeDirected Molecular EvolutionAspartic AcidDNAAmino Acid MotifsEnzyme StabilityMutant ProteinsPromoter Regions, GeneticUltraviolet RaysSaccharomyces cerevisiaeTranscription, GeneticAsparagineCricetinaeGene Expression Regulation, BacterialGenetic VectorsDNA-Binding ProteinsSerineCOS CellsPolymerase Chain ReactionDNA RepairDNA-Directed DNA PolymeraseArginineCircular DichroismTranscription FactorsSequence Analysis, DNADimerizationLigandsTryptophanGene DeletionCHO CellsCercopithecus aethiopsCodonElectrophoresis, Polyacrylamide GelMutagenicity TestsAmino AcidsOperonHydrogen-Ion ConcentrationGlycosylationGene LibraryMembrane ProteinsRecombination, GeneticCarrier ProteinsDNA DamageTransformation, BacterialHydrolysisTransformation, GeneticMethylnitronitrosoguanidineGenetic EngineeringHydrogen Bonding
Risk of insertional mutagenesis comparedTransgenesis and insertional mutagenesisVectorsGenomeSite-directed muTransposonRetrovirusesGenesGene therapyMammalianScreensTransformationIntegrationInsertionCas9GeneticSequencesExpressionSequenceCellRisk of insertional mutagenesisSomaticMutationsRetroviralCharacterizationCRISPRDisruptionTranspositionGenerateLymphomaTherapeuticMiceToolHostTol2Cell lineCellsLarge-scaleActivationMethodMouseProcessCancer
Risk of insertional mutagenesis compared1
  • Theoretically, this approach decreases the risk of insertional mutagenesis compared with repeated, separate integrations of multiple genes and associated promoters into the T-cell genome. (ubigene.us)
Transgenesis and insertional mutagenesis2
  • These integration systems are ideally suited to transgenesis and insertional mutagenesis strategies in the frog. (biomedcentral.com)
  • Since the first discovery of DNA transposons in Maize by Barbara#McClintock in 1950 [ 1 ], transposons have been used extensively as genetic tools in invertebrates and in plants for transgenesis and insertional mutagenesis [ 2 - 7 ]. (biomedcentral.com)
Vectors9
  • Transposon vectors can be used in the frog for transgenesis and for insertional mutagenesis where enhancer trap and gene trap constructs are used to identify genomic loci involved in developmental processes. (biomedcentral.com)
  • Viral vectors also have the largest genomic footprint of iPSC generating methods due to the risk of insertional mutagenesis. (addgene.org)
  • Gene delivery strategies based on viral vectors are currently the most efficient. (tamu.edu)
  • Despite the broad tropism of VSV-G, lentivirus vector transduction is less efficient than adenoviral vectors. (alecomm.com)
  • Third generation lentiviral vectors currently in use encode no viral proteins, only a few cis-acting sequence elements required for packaging, reverse transcription, and efficient expression. (alecomm.com)
  • It has been speculated that accidental exposure to lentiviral vectors could result in insertional mutagenesis, a fear found on observed tendency for the closely related γ-retroviral vectors to integrate in proximity to proto-oncogenes. (alecomm.com)
  • Extensive analysis has shown though that lentiviral vectors have a very different preference for insertion sites, and so hazardous insertional mutagenesis is unlikely. (alecomm.com)
  • To achieve efficient genetic material delivery in mammalian cells, viral-derived vectors have been served as the most promising tools. (creative-biolabs.com)
  • In this important study, Drs. Thomas and Capecchi used both replacement and insertional vectors to introduce a neomycin resistance (neo r ) gene into exon 8 of the Hprt gene, showing that clones of transfected cells had lost HPRT function (becoming 6-TG resistant), and had gained Neo r activity. (ijbs.com)
Genome4
  • Until recently, however, functional analysis of the DED fungi was hampered by the lack of protocols for efficient gene knockout and by the unavailability of a full genome sequence. (sisef.it)
  • The high fecundity of the frog combined with the ability to remobilize transposon transgenes integrated into frog genome will allow large-scale insertional mutagenesis screens to be performed in laboratories with modest husbandry capacities. (biomedcentral.com)
  • Dr. Burgess is now merging large-scale mutagenesis and phenotype characterization into a novel approach to functionally annotate a vertebrate genome. (nih.gov)
  • Because of their ability to generate insertions, IS elements and transposons represent a useful and efficient tool in biotechnology by introducing 'foreign' DNA into the genome of various plants, animals or bacteria (for a review, see Coates et al. (proteasomesignaling.com)
Site-directed mu1
Transposon2
  • Insertional transposon mutagenesis is a frequently used technique with the enormous advantage not only of the generation of new phenotypes, but the identification of the mutated gene directly. (proteasomesignaling.com)
  • Transposon mutagenesis can be achieved by several means including both random and site-directed methods. (proteasomesignaling.com)
Retroviruses2
  • Finally, retroviruses are prone to insertional mutagenesis and random integration creates variation between iPSC clones. (addgene.org)
  • tax causes malignant transformation not through integration and insertional mutagenesis, as many retroviruses do, but by inhibition of DNA repair (base excision pathway) and trans-activating disruption of cellular growth control mechanisms ( 2 ). (cdc.gov)
Genes6
  • While the selective inactivation of Ophiostoma genes by insertional mutagenesis remains a challenge, an alternative approach based on RNA interference is now available for down-regulating the expression of targeted genes. (sisef.it)
  • Efficient integration of functional genes is an essential prerequisite for successful gene delivery such as cell transfection, animal transgenesis, and gene therapy. (tamu.edu)
  • As a complementary resource, chemically induced mutants have been shown to provide an efficient alternative because each individual line can bear single point missense and nonsense substitutions in hundreds of genes [ 7 ]. (biomedcentral.com)
  • More recently, Dr. Burgess has turned his mutagenesis efforts to targeted knockouts using CRISPR/ Cas9 technology, gaining full control over which genes are inactivated. (nih.gov)
  • MMTV causes tumors by insertional mutagenesis, i.e., viral DNA integration near Wnt or Fgf genes, for breast cancer induction. (utexas.edu)
  • Multiple mutations affecting embryonic development have been induced by chemical and insertional mutagenesis resulting in the identification of genes with important roles in development in Drosophila [ 23 - 25 ]. (biomedcentral.com)
Gene therapy3
  • In recent years, these powerful genetic tools have been used in higher vertebrates for transgenesis, insertional mutagenesis, and gene therapy applications. (biomedcentral.com)
  • The results presented herein demonstrate that our helper-independent plasmids represent an important step in the development of safe and efficient gene delivery methods that should prove valuable in gene therapy and transgenic approaches. (tamu.edu)
  • DNA transposons have emerged as indispensible tools for manipulating vertebrate genomes with applications ranging from insertional mutagenesis and transgenesis to gene therapy. (biomedcentral.com)
Mammalian1
  • This work led to his important conclusion that mammalian somatic cells possess an efficient enzymatic machinery for mediating homologous recombination [ 6 ]. (ijbs.com)
Screens2
  • The rapid accumulation of genetic resources for this animal and the success of pilot mutagenesis screens have helped propel this model system forward. (biomedcentral.com)
  • Knowledge of the genetic networks controlling embryogenesis has been obtained principally by mutagenesis screens in model species. (biomedcentral.com)
Transformation1
Integration1
Insertion1
  • Site-directed or targeted mutagenesis mediated by insertion sequence (IS) elements and transposons relates to the use of a novel recombinant DNA technology for the targeted modification of DNA. (proteasomesignaling.com)
Cas91
  • In this report, we described the application of CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (endonuclease) system for efficient genetic editing in E. tenella , and showed that the CRISPR/Cas9 system mediates site-specific double-strand DNA breaks with a single guide RNA. (biomedcentral.com)
Genetic2
  • Due to lack of efficient in vitro tissue culture model and an effective selection marker, the technical capability for genetic modification of Eimeria has lagged behind for that of Toxoplasma and Plasmodium . (biomedcentral.com)
  • Dr. Burgess' mutagenesis and phenotyping efforts address a variety of questions ranging from systematically characterizing cell signaling molecules to generating animal models for rare genetic diseases. (nih.gov)
Sequences1
  • Over several serial transfers, particular sequences accumulated, and in isolation these were shown to out-compete less efficient replicators. (korea.ac.kr)
Expression2
  • The enhancers, along with the LTR promoter sequence, supported efficient expression of the corrective IL2RG gene and allowed successful gene correction. (europeanhealthjournal.com)
  • Transcriptomic analyses provide an efficient approach to explore the temporal gene expression profiles in embryos and to search for the developmental regulators. (biomedcentral.com)
Sequence1
  • He reasoned that if this machinery could be efficient for homologous recombination between a newly introduced DNA molecule and the endogenous DNA sequence, any host gene could be mutated. (ijbs.com)
Cell2
  • At the subcellular level, CD19/20/22CAR T-cells formed dense immune synapses with target cells that mediated effective cytolytic complex formation, were efficient serial killers in single-cell tracking studies, and were as efficacious as CD19CAR T-cells against primary CD19(+) disease. (ubigene.us)
  • In their paper, they concluded 'It is hoped that this combination of using ES cells as the recipient cell line and site-specific mutagenesis achieved by gene targeting will provide the means for generating mice of any desired genotype" [ 2 ]. (ijbs.com)
Risk of insertional mutagenesis3
  • As they remain episomal they have minimal-to-no risk of insertional mutagenesis. (medscape.com)
  • This uncontrollable integration generates risk of insertional mutagenesis, which can produce undesirable effects on the modified cells. (ugr.es)
  • However, as Signal Transduction and Targeted Therapy notes, LV vectors have historically carried the risk of insertional mutagenesis, though this risk can be minimized by using third-generation, self-inactivating LV vectors. (corning.com)
Somatic2
  • 1. Somatic mutagenesis with a Sleeping Beauty transposon system leads to solid tumor formation in zebrafish. (nih.gov)
  • CRISPR/Cas9 somatic multiplex-mutagenesis for high-throughput functional cancer genomics in mice. (nature.com)
Mutations4
  • This RFA provides a flexible system within the SBIR/STTR programs to cover the research steps needed to develop and validate technology to generate insertional mutations in mouse ES cells. (nih.gov)
  • ii) The study of re- troviruses lacking their own oncogenes provided the first direct evidence that known cellular oncogenes could participate in carcinogenesis, as the targets for activation by viral insertional mutations. (nih.gov)
  • Most ERVs in each class are no longer capable of transcription and/or retrotransposition due to the accumulation of mutations and/or efficient targeting by host silencing mechanisms that act at various stages of the viral life cycle [ 2 ]. (biomedcentral.com)
  • The main drawback of these technologies is the relatively low frequency of mutations, as compared to those induced by conventional chemical or physical agents, thus limiting the use of insertional mutagens to the generation of large mutant populations in few genetic backgrounds. (ncsu.edu)
Retroviral1
  • We introduce a multi-scale diffusion kernel and apply it to a large collection of murine retroviral insertional mutagenesis data. (dtls.nl)
Characterization3
  • Interests include, but are not limited to, development of new methods for mutagenesis and/or phenotypic characterization that would be of use in research on a wide range of diseases or organs, particularly if these methods could be applied to other animal models as well as the zebrafish. (nih.gov)
  • Dr. Burgess is now merging large-scale mutagenesis and phenotype characterization into a novel approach to functionally annotate a vertebrate genome. (nih.gov)
  • Our genetic and molecular characterization of slvipp1 , an insertional mutant, allowed us to conclude that the tomato SlVIPP1 gene is needed for development, as Arabidopsis VIPP1 , but not Escherichia coli PspA . (frontiersin.org)
CRISPR2
  • The strategy utilizes TIM, a CRISPR/Cas9-based method for targeted insertional mutagenesis in C. reinhardtii. (umassmed.edu)
  • More recently, Dr. Burgess has turned his mutagenesis efforts to targeted knockouts using CRISPR/ Cas9 technology, gaining full control over which genes are inactivated. (nih.gov)
Disruption2
  • The SFB1530 brings together the synergistic combination of efficient disruption of lymphoma cell-intrinsic oncogenic programs and a specific modulation of the lymphoma (immune) microenvironment. (sfb1530.de)
  • tax causes malignant transformation not through integration and insertional mutagenesis, as many retroviruses do, but by inhibition of DNA repair (base excision pathway) and trans-activating disruption of cellular growth control mechanisms ( 2 ). (cdc.gov)
Transposition2
  • 8. Efficient transposition of the Tol2 transposable element from a single-copy donor in zebrafish. (nih.gov)
  • MuDR exhibits the highest transposition activity and insertional mutagenesis frequency in Mutator (Mu) family. (sachecucine.it)
Generate1
  • Herein, we describe a simple yet robust Arabidopsis transformation procedure that can be used to generate large numbers of insertional mutants in Arabidopsis thaliana. (ncsu.edu)
Lymphoma1
  • 18. Whole-body sleeping beauty mutagenesis can cause penetrant leukemia/lymphoma and rare high-grade glioma without associated embryonic lethality. (nih.gov)
Therapeutic1
  • The efficiency, safety and cost of this process need to be optimized to ensure therapeutic efficacy, to mitigate potential adverse events such as insertional mutagenesis and to improve the cost-effectiveness of gene therapies. (journalforclinicalstudies.com)
Mice1
  • 15. PiggyBac transposon mutagenesis: a tool for cancer gene discovery in mice. (nih.gov)
Tool2
  • This effective, efficient, and convenient TIM-tagging strategy promises to be a useful tool for the study of nuclear genes, including essential genes, in C. reinhardtii. (umassmed.edu)
  • Our MSCV retrovirus CAR expression vector is a highly efficient tool for retrovirus-based delivery of second-generation CAR expression cassettes into T cells. (vectorbuilder.com)
Host1
  • Taken together, our data suggest that MoHDA1-dependent histone deacetylation is important for efficient asexual development and infection of host plants in M. oryzae . (ppjonline.org)
Tol21
  • piggy Bac (PB), one of three transposons (PB, Sleeping Beauty (SB), and Tol2 ) found thus far, has been used for efficient transfection of GOI in various mammalian cells in vitro and in vivo. (encyclopedia.pub)
Cell line1
  • Markowitz, D., Goff, S. & Bank, A. Construction and use of a safe and efficient amphotropic packaging cell line. (swallowxx.com)
Cells3
  • Ads are efficient in infecting a wide variety of cell types and exhibiting high-level transient transgene expression in target cells. (medscape.com)
  • Since membranes play essential roles in all living beings, all cells have developed mechanisms for efficient and fast repair of membrane damage. (frontiersin.org)
  • The most direct and efficient approach for cloning parasitism genes and rapidly advancing our understanding of the molecular interactions during nematode parasitism of plants is to create gland cell-specific cDNA libraries using cytoplasm microaspirated from the esophageal gland cells of various parasitic stages. (ncsu.edu)
Large-scale1
  • Uniquely for emerging model organisms, T. castaneum offers the opportunity of performing time- and cost-efficient large-scale RNAi screening, based on commercially available dsRNAs targeting all genes, which are simply injected into the body cavity. (biomedcentral.com)
Activation1
  • the Wnt-1 gene, discovered as a target for insertional activation by the mouse mammary tumor virus, belongs to a large family of genes encoding secretory proteins involved in important developmental events in many organisms. (nih.gov)
Method1
  • The experimental results demonstrated that CDL-PCR is an efficient and flexible method for identifying the flanking sequences in transgenic rice and Arabidopsis …In this protein family, a conserved core domain, the so-called α-crystallin or Hsp20 domain, is flanked by highly variable, nonconserved sequences that are essential for chaperone function. (sachecucine.it)
Mouse1
  • This RFA, "Tools for Insertional Mutagenesis in the Mouse: SBIR/STTR Initiative," is related to several of the priority areas. (nih.gov)
Process1
  • Oxygen production is a crucial and highly efficient process that uses solar energy to convert carbon dioxide and water into sugars and other organic compounds, hence providing resources for plant and animal life. (frontiersin.org)
Cancer1