SpringerProtocols: Abstract: 12. Using Nucleases to Stimulate Homologous Recombination

12. Using Nucleases to Stimulate Homologous Recombination

By: Dana Carroll²

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In essentially all organisms, double-strand breaks in chromosomal DNA stimulate repair by multiple mechanisms, including homologous recombination. It is possible to use site-specific reagents to produce a break or other recombinagenic damage at a unique site, which makes possible detailed analysis of the repair products. In addition, targeted mutagenesis and gene replacement are stimulated in the immediate vicinity of the break site. To utilize meganucleases with long recognition sequences, it is necessary to introduce the corresponding sequence prior to directed cleavage. The same is typically true of triplex-forming oligonucleotides that target polypurine-polypyrimidine tracts. Zinc-finger nucleases have the potential of being targetable to arbitrarily selected sites, owing to the flexibility of zinc finger recognition of DNA.

Affiliation(s): (2) Department of Biochemistry II, University of Utah School of Medicine, Salt Lake City, UT

Book Title: Genetic Recombination: Reviews and Protocols

Series: Methods in Molecular Biology | Volume: 262 | Pub. Date: Jan-09-2004 | Page Range: 195-207 | DOI: 10.1385/1-59259-761-0:195

Subject: Genetics/Genomics

Key Words: genetic recombination - homologous recombination - nonhomologous end joining - I-SceI - zinc fingers - zinc finger nucleases - double-strand break - DNA repair - gene targeting

References

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