Research ArticleGenome Editing

Efficient modification of CCR5 in primary human hematopoietic cells using a megaTAL nuclease and AAV donor template

See allHide authors and affiliations

Science Translational Medicine  30 Sep 2015:
Vol. 7, Issue 307, pp. 307ra156
DOI: 10.1126/scitranslmed.aac5530

Delete and replace

Newer gene-editing methods hold promise for correcting human disease but so far have been hampered by low efficiencies when used in primary cells. To address this issue, Sather et al. have devised a more effective way to both disrupt and replace the CCR5 locus in human T cells, a procedure that has already been shown to improve HIV clearance.

Serotype 6 of an adeno-associated viral vector worked particularly well for delivery of megaTAL nucleases and homologous donor templates to primary human T cells, achieving efficient gene-editing rates and little toxicity. The megaTALs generate homology-directed repair (rather than previous efforts, which induce nonhomologous end-joining repair) and so was used for both deletion and accurate replacement of the CCR5 locus. The authors demonstrate that chimeric antigen receptors and an HIV fusion inhibitor inserted into the CCR5 locus ameliorate HIV infection in mice and show that their approach also works in CD34+ hematopoietic precursor cells.

View Full Text

Stay Connected to Science Translational Medicine