ArticleMolecular biotechnology2009
Increased gene targeting in Ku70 and Xrcc4 transiently deficient human somatic cells.
Article in Molecular biotechnology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Cellular localization of a variant RAPGEF5 protein associated with idiopathic epilepsy risk in the Belgian shepherd.Canine medicine and genetics · 2024Article
- Generation and validation of homozygous fluorescent knock-in cells using CRISPR-Cas9 genome editing.Nature protocols · 2018Article
- Biallelic Gene Targeting in Rice.Plant physiology · 2016Article
- Targeting non-coding RNAs with the CRISPR/Cas9 system in human cell lines.Nucleic acids research · 2015Article
- Improved Gene Targeting through Cell Cycle Synchronization.PloS one · 2015Article
- Aptamer-guided gene targeting in yeast and human cells.Nucleic acids research · 2014Article
- CRISPR/Cas9 mediated multiplex genome editing and heritable mutagenesis of BmKu70 in Bombyx mori.Scientific reports · 2014Article
- Recent advances in the development of new transgenic animal technology.Cellular and molecular life sciences : CMLS · 2013Review
- Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice.The New phytologist · 2012Article
- Agrobacterium may delay plant nonhomologous end-joining DNA repair via XRCC4 to favor T-DNA integration.The Plant cell · 2012Article
- Chemical and biological approaches to improve the efficiency of homologous recombination in human cells mediated by artificial restriction DNA cutter.Nucleic acids research · 2012Article
- Efficient and accurate homologous recombination in hESCs and hiPSCs using helper-dependent adenoviral vectors.Molecular therapy : the journal of the American Society of Gene Therapy · 2012Article
- An update on targeted gene repair in mammalian cells: methods and mechanisms.Journal of biomedical science · 2011Review
- Identification of genes regulating gene targeting by a high-throughput screening approach.Journal of nucleic acids · 2011Article
- Transgenic pigs as models for translational biomedical research.Journal of molecular medicine (Berlin, Germany) · 2010Review
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The insertion of foreign DNA at a specific genomic locus directed by homologous DNA sequences, or gene targeting, is an inefficient process in mammalian somatic cells. Given the key role of non-homologous end joining (NHEJ) pathway in DNA double-strand break (DSB) repair in mammalian cells, we investigated the effects of decreasing NHEJ protein levels on gene targeting. Here we demonstrate that the transient knockdown of integral NHEJ proteins, Ku70 and Xrcc4, by RNAi in human HCT116 cells has a remarkable effect on gene targeting/random insertions ratios. A timely transfection of an HPRT-based targeting vector after RNAi treatment led to a 70% reduction in random integration events and a 33-fold increase in gene targeting at the HPRT locus. These findings bolster the role of NHEJ proteins in foreign DNA integration in vivo, and demonstrate that their transient depletion by RNAi is a viable approach to increase the frequency of gene targeting events. Understanding how foreign DNA integrates into a cell's genome is important to advance strategies for biotechnology and genetic medicine.
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Registered trials
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