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  • Cas9
  • (pnas.org)
  • These studies should greatly accelerate investigations into mosquito biology, streamline development of transgenic strains for field releases, and simplify the evaluation of novel Cas9-based gene drive systems. (pnas.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)
  • When gRNA s targeting ALS were transcribed transiently from double-stranded T-DNA containing the HDR template, DSB s were induced in the ALS locus by the assembled Cas9 / gRNA complex and homologous recombination was stimulated. (plantphysiol.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)
  • knockout
  • Unexpectedly, studies involving primarily knockout mice also implicated MMR genes in other processes of DNA metabolism, ranging from DNA damage signaling to hypermutation of Ig genes, where the molecular roles of MMR proteins are unclear ( 1 ). (pnas.org)
  • locus
  • The injection procedure is much more rapid than earlier approaches and makes possible the generation and recovery of targeted gene alterations at essentially any locus within 2 fly generations. (pnas.org)
  • In these experiments, which were performed in primary cells and in established cell lines, the frequency of homologous recombination of the HC-AdV DNA with the HPRT locus was found to be, depending on the cell type, between 2x10-5 to 1.2x10-6. (uni-koeln.de)
  • We now describe the generation of chicken DT40 B cells, in which the FAN1 locus was disrupted by gene targeting. (pnas.org)
  • cells
  • H3-G34R mutants are defective for DNA damage repair by homologous recombination (HR), and have altered HR protein dynamics in both damaged and untreated cells. (elifesciences.org)
  • They help to protect the DNA from damage, control when genes are switched on or off, and influence how the DNA is copied when cells are preparing to divide to produce two daughter cells. (elifesciences.org)
  • In order to understand regulation of both host and virus gene expression, we have utilized genome-wide approaches to analyze transcript abundance from both virus and host cells. (jove.com)
  • Cells also repair DSBs by homologous recombination (HR), and modified versions of the target gene supplied by the experimenter can serve as the repair template, thereby introducing designed modifications ( 10 , 11 ). (pnas.org)
  • The cells do not display elevated sister chromatid exchange frequencies, either sporadic or MMC-induced. (pnas.org)
  • Despite success with the technique, recent studies have revealed that following repair events, correction frequencies decrease as a function of time, possibly due to a sustained activation of damage response signals in corrected cells that lead to a selective stalling. (biomedcentral.com)
  • Taken together, we believe that thymidine may be used in a therapeutic fashion to enable the maintenance of high levels of treated cells bearing repaired genes. (biomedcentral.com)
  • We demonstrate that guide RNA pairs generate deletions that are repaired with a high level of precision by non-homologous end-joining in mammalian cells. (stanford.edu)
  • These cells are made when cells containing two copies of every gene in the organism divide to produce new cells that each only have one copy of each gene. (elifesciences.org)
  • The poison is capable of killing all gametes, but the antidote protects the cells that contain the gamete killer gene. (elifesciences.org)
  • Further experiments show that the antidote produced by one of the gamete killer genes cannot protect cells against the poison produced by the other gene. (elifesciences.org)
  • During this time his work has been the first to demonstrate that gene correction could be achieved in human cells at frequencies that were high enough to potentially cure patients and is considered one of the pioneers and founders of the field of genome editing-a field that now encompasses thousands of labs and several new companies throughout the world. (stanford.edu)
  • Individuals with Primary Immune Deficiency (PID) may develop severe, life-threatening infections as a result of inherited defects in the genes that normally instruct blood-forming cells to develop and to fight infections. (stanford.edu)
  • The reported phenotype provides the first genetic evidence of a link between double-strand break repair and homologous recombination in vertebrate cells. (nih.gov)
  • 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)
  • Making targeted gene replacements has also become standard practice in mice, thanks to the availability of embryonic stem (ES) cells that can be manipulated in culture and the development of powerful selection procedures ( Capecchi 2005 ). (genetics.org)
  • With other experimental organisms, ES cells are not available, screening or selection procedures are not adequate, and development of useful gene-targeting approaches is impeded by the low frequency of recombination. (genetics.org)
  • Through recombination, daughter cells have the greatest amount of genetic diversity. (wikipedia.org)
  • encodes
  • Many of these lesions cause structural damage to the DNA molecule and can alter or eliminate the cell's ability to transcribe the gene that the affected DNA encodes. (wn.com)
  • phenotype
  • Explain how continuous traits are the result of many different gene combinations that can each contribute a varying amount to a phenotype. (genetics-gsa.org)
  • Evaluate how genes and the environment can interact to produce a phenotype. (genetics-gsa.org)
  • maternal
  • Mendel's law of equal segregation stipulates that paternal and maternal alleles of a gene should have an equal chance of being transmitted to progenies. (elifesciences.org)
  • pombe
  • We propose that wtf genes contribute to the extensive intraspecific reproductive isolation in S. pombe , and represent ideal models for understanding how segregation-distorting elements act and evolve. (elifesciences.org)
  • vector
  • About half of the vector DNA integrations took place inside genes and none were observed in protooncogenes. (uni-koeln.de)
  • mediate
  • While several genes essential for bacterial cytokinesis have been identified ( 2 , 3 ), no gene product has been found to mediate organelle division in a eukaryote. (pnas.org)
  • protein
  • For example, a large, complete deletion of a protein coding gene will obviously lead to the elimination of the expression of that protein. (biomedcentral.com)
  • ZFNs
  • We report very high gene targeting frequencies in Drosophila by direct embryo injection of mRNAs encoding specific zinc-finger nucleases (ZFNs). (pnas.org)
  • The procedure that generates these high frequencies relies on cleavage of the target by designed zinc-finger nucleases (ZFNs) and production of a linear donor in situ . (genetics.org)
  • The bright prospects for future applications of ZFNs, including human gene therapy, are discussed. (genetics.org)
  • events
  • For DSB-induced events, similar recombination frequencies and conversion tract spectra were found under conditions of low and high transcription. (asm.org)
  • We describe methods for detecting homologous recombination (HR) events, including direct screening methods as well as new selection/counterselection strategies. (genetics.org)
  • A transformation scheme for Cryptococcus neoformans to yield high-frequency, integrative events was developed. (asm.org)
  • Overall, our results support the emerging notion that metabolizing gene families, such as the GSTM, NAT, UGT and CYP , have been evolving rapidly through gene duplication and deletion events in primates, leading to complex structural variation within and among species with unknown evolutionary consequences. (biomedcentral.com)
  • If linkage is complete, there should be no recombination events that separate the two alleles, and therefore only parental combinations of alleles should be observed in offspring. (wikipedia.org)
  • nucleases
  • RNAi-based suppression of Ku70 concurrent with embryonic microinjection of site-specific nucleases yielded consistent gene insertion frequencies of 2-3%, similar to traditional transposon- or ΦC31-based integration methods but without the requirement for an initial docking step. (pnas.org)
  • approaches
  • Our research is focused on development of novel strategies for gene and cell therapy, using gene therapy and regenerative medicine approaches. (stanford.edu)
  • integration
  • A RAD54 homolog was disrupted in the chicken B cell line DT40, which undergoes immunoglobulin gene conversion and exhibits unusually high ratios of targeted to random integration after DNA transfection. (nih.gov)
  • eukaryotes
  • In eukaryotes, the recombinase RAD51 forms a nucleoprotein filament on ssDNA that performs synapsis and strand invasion of the homologous duplex DNA to form a stable D-loop. (elifesciences.org)
  • therapies
  • In this review, we discuss the roles the Mycs play in the body and what led us to choose them to be our candidate gene for inner ear therapies. (mdpi.com)
  • Cell, tissue, and gene therapies. (stanford.edu)
  • tandem
  • Finally, we used a collection of tandem repeats (from 16 to 935 bp in length) within the mtr gene to examine the length requirement for RIP. (genetics.org)
  • Mechanisms for altering or generating tandem gene arrays. (g3journal.org)
  • high
  • Our findings reveal a surprisingly high frequency of HR-mediated gene conversion, making it possible to rapidly and precisely edit the C. elegans genome both with and without the use of co-inserted marker genes. (genetics.org)