title: CRISPR/Cas9-Derived Mutations Both Inhibit HIV-1 Replication and Accelerate Viral Escape.
abstract: Cas9 cleaves specific DNA sequences with the assistance of a programmable single guide RNA (sgRNA). Repairing this broken DNA by the cell's error-prone non-homologous end joining (NHEJ) machinery leads to insertions and deletions (indels) that often impair DNA function. Using HIV-1, we have now demonstrated that many of these indels are indeed lethal for the virus, but that others lead to the emergence of replication competent viruses that are resistant to Cas9/sgRNA. This unexpected contribution of Cas9 to the development of viral resistance is facilitated by some indels that are not deleterious for viral replication, but that are refractory to recognition by the same sgRNA as a result of changing the target DNA sequences. This observation illustrates two opposite outcomes of Cas9/sgRNA action, i.e., inactivation of HIV-1 and acceleration of viral escape, thereby potentially limiting the use of Cas9/sgRNA in HIV-1 therapy.
authors.affiliation: McGill University AIDS Centre, Lady Davis Institute, Jewish General Hospital, Montreal H3T 1E2, Canada; Departments of Medicine, Microbiology and Immunology, McGill University, Montreal H3A 2B4, Canada.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.affiliation: McGill University AIDS Centre, Lady Davis Institute, Jewish General Hospital, Montreal H3T 1E2, Canada.
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Pan
authors.affiliation: Institute for Research in Immunology and Cancer, University of Montreal, Montreal H3C 3J7, Canada.
authors.forename: Patrick
authors.initials: P
authors.lastname: Gendron
authors.affiliation: MOH Key Laboratory of Systems Biology of Pathogens and Center for AIDS Research, Institute of Pathogen Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China.
authors.forename: Weijun
authors.initials: W
authors.lastname: Zhu
authors.affiliation: MOH Key Laboratory of Systems Biology of Pathogens and Center for AIDS Research, Institute of Pathogen Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Guo
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing 100050, China.
authors.forename: Shan
authors.initials: S
authors.lastname: Cen
authors.affiliation: McGill University AIDS Centre, Lady Davis Institute, Jewish General Hospital, Montreal H3T 1E2, Canada; Departments of Medicine, Microbiology and Immunology, McGill University, Montreal H3A 2B4, Canada.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Wainberg
authors.affiliation: McGill University AIDS Centre, Lady Davis Institute, Jewish General Hospital, Montreal H3T 1E2, Canada; Departments of Medicine, Microbiology and Immunology, McGill University, Montreal H3A 2B4, Canada. Electronic address: chen.liang@mcgill.ca.
authors.forename: Chen
authors.initials: C
authors.lastname: Liang
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D000072669:Gene Editing; D015497:HIV-1 / Q000235:genetics* / Q000254:growth & development / Q000502:physiology*; D006801:Humans; D054643:INDEL Mutation / Q000235:genetics; D057131:Immune Evasion; D009154:Mutation / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
chemical_list: D004279:DNA, Viral