Targeted disruption of the CCR5 gene in human hematopoietic stem cells stimulated by peptide nucleic acids.
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
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.
Schleifman, Erica B; Bindra, Ranjit; Leif, Jean; del Campo, Jacob; Rogers, Faye A; Uchil, Pradeep; Kutsch, Olaf; Shultz, Leonard D; Kumar, Priti; Greiner, Dale L; and Glazer, Peter M, "Targeted disruption of the CCR5 gene in human hematopoietic stem cells stimulated by peptide nucleic acids." (2011). Faculty Research 2011. 167.