RT Journal
A1 Kim, Hye Joo
A1 Lee, Hyung Joo
A1 Kim, Hyojin
A1 Cho, Seung Woo
A1 Kim, Jin-Soo
T1 Targeted genome editing in human cells with zinc finger nucleases constructed via modular assembly
JF Genome Research 
JO Genome Research 
YR 2009 
FD July 01 
VO 19 
IS 7 
SP 1279 
OP 1288 
DO 10.1101/gr.089417.108 
UL http://genome.cshlp.org/content/19/7/1279.abstract 
AB 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.