ArticleHuman gene therapy2008
Human gene targeting favors insertions over deletions.
Article in Human gene therapy, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- CRISPR Genome Engineering for Human Pluripotent Stem Cell Research.Theranostics · 2017Review
- Adeno-associated Virus as a Mammalian DNA Vector.Microbiology spectrum · 2015Review
- A genome-wide map of adeno-associated virus-mediated human gene targeting.Nature structural & molecular biology · 2014Article
- Sgs1 and Exo1 suppress targeted chromosome duplication during ends-in and ends-out gene targeting.DNA repair · 2014Article
- The effects of polymorphisms on human gene targeting.Nucleic acids research · 2014Article
- Targeted gene addition to a predetermined site in the human genome using a ZFN-based nicking enzyme.Genome research · 2012Article
- Directed evolution of adeno-associated virus for enhanced gene delivery and gene targeting in human pluripotent stem cells.Molecular therapy : the journal of the American Society of Gene Therapy · 2012Article
- Article
- AAV-mediated gene targeting methods for human cells.Nature protocols · 2011Article
- Single-strand nicks induce homologous recombination with less toxicity than double-strand breaks using an AAV vector template.Nucleic acids research · 2011Article
- Efficient KRT14 targeting and functional characterization of transplanted human keratinocytes for the treatment of epidermolysis bullosa simplex.Molecular therapy : the journal of the American Society of Gene Therapy · 2010Article
- Chromosomal position effects on AAV-mediated gene targeting.Nucleic acids research · 2010Article
- Engineering of human pluripotent stem cells by AAV-mediated gene targeting.Molecular therapy : the journal of the American Society of Gene Therapy · 2010Article
Corrections and comments
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Gene targeting is a powerful technique for manipulating the human genome, but few studies have directly compared the targeting frequencies of various types of vector constructs. Here we show that similar targeting constructs are able to insert nucleotides at the homologous chromosomal target locus more efficiently than they can delete nucleotides, and combination insertion/deletion vectors appear to target at intermediate frequencies. This holds true for deletions ranging from 1 to 334 bp and insertions ranging from 1 to 1332 bp. In addition, vectors designed to inactivate the human hypoxanthine phosphoribosyltransferase gene (HPRT) by deleting nucleotides often produced rearrangements at the target locus that in many cases were due to insertions of multimerized vector constructs, effectively converting a deletion vector into an insertion vector. These findings were obtained when adeno-associated virus vectors were used to efficiently deliver single-stranded DNA targeting constructs, but the same phenomenon was also observed when transfecting linearized double-stranded plasmids. Thus human cells distinguish between deletion and insertion vectors and process their recombination intermediates differently, presumably at the heteroduplex stage, with implications for the design of gene-targeting vectors and the evolution of human genomes.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.