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PMID: 19470664 Published · ppublish English Journal Article

Targeted genome editing in human cells with zinc finger nucleases constructed via modular assembly.

Genome research ·Vol. 19 ·No. 7 ·2009-07-00 ·Pages 1279-88

Kim HJ, Lee HJ, Kim H, Cho SW, Kim JS

Abstract

Broad applications of zinc finger nuclease (ZFN) technology-which allows targeted genome editing-in research, medicine, and biotechnology are hampered by the lack of a convenient, rapid, and publicly available method for the synthesis of functional ZFNs. Here we describe an efficient and easy-to-practice modular-assembly method using publicly available zinc fingers to make ZFNs that can modify the DNA sequences of predetermined genomic sites in human cells. We synthesized and tested hundreds of ZFNs to target dozens of different sites in the human CCR5 gene-a co-receptor required for HIV infection-and found that many of these nucleases induced site-specific mutations in the CCR5 sequence. Because human cells that harbor CCR5 null mutations are functional and normal, these ZFNs might be used for (1) knocking out CCR5 to produce T-cells that are resistant to HIV infection in AIDS patients or (2) inserting therapeutic genes at "safe sites" in gene therapy applications.

MeSH Terms
Base Sequence DNA Breaks, Double-Stranded Endonucleases/genetics,metabolism Genome, Human Humans Kidney/cytology,metabolism Luciferases/metabolism Molecular Sequence Data Mutation Receptors, CCR5/genetics,metabolism Sequence Homology, Nucleic Acid Zinc Fingers/genetics
Chemicals
Receptors, CCR5 Luciferases Endonucleases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Kim Hye Joo
Department of Chemistry, Seoul National University, Gwanak-gu, Seoul 151-742, South Korea.
Lee Hyung Joo
Kim Hyojin
Cho Seung Woo
Kim Jin-Soo
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Article Info
Journal
Genome research
Abbr.
Genome Res
ISSN
1088-9051
DOI
10.1101/gr.089417.108
Published
2009-07-00
Epub
2009-00-21
Pages
1279-88
Language
English
Region
United States
NLM ID
9518021
PMCID
PMC2704428
Subset
IM
External Links
PubMed source DOI: 10.1101/gr.089417.108 PMC Full Text View journal details
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