Targeted disruption of the CCR5 gene in human hematopoietic stem cells stimulated by peptide nucleic acids.

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Publication Date



Animals, Antigens, CD34, Base Sequence, Binding Sites, Cell Line, Codon, Terminator, DNA Repair, Gene Targeting, HIV Infections, HIV-1, Hematopoietic Stem Cells, Humans, Mice, Mutation, Peptide Nucleic Acids, Receptors, CCR5

JAX Source

Chem Biol 2011 Sep 23; 18(9):1189-98.




Peptide nucleic acids (PNAs) bind duplex DNA in a sequence-specific manner, creating triplex structures that can provoke DNA repair and produce genome modification. CCR5 encodes a chemokine receptor required for HIV-1 entry into human cells, and individuals carrying mutations in this gene are resistant to HIV-1 infection. Transfection of human cells with PNAs targeted to the CCR5 gene, plus donor DNAs designed to introduce stop codons mimicking the naturally occurring CCR5-delta32 mutation, produced 2.46% targeted gene modification. CCR5 modification was confirmed at the DNA, RNA, and protein levels and was shown to confer resistance to infection with HIV-1. Targeting of CCR5 was achieved in human CD34(+) hematopoietic stem cells (HSCs) with subsequent engraftment into mice and persistence of the gene modification more than four months posttransplantation. This work suggests a therapeutic strategy for CCR5 knockout in HSCs from HIV-1-infected individuals, rendering cells resistant to HIV-1 and preserving immune system function.