A combinational CRISPR/Cas9 gene-editing approach can halt HIV replication and prevent viral escape.
HIV presents one of the highest evolutionary rates ever detected and combination antiretroviral therapy is needed to overcome the plasticity of the virus population and control viral replication. Conventional treatments lack the ability to clear the latent reservoir, which remains the major obstacle towards a cure. Novel strategies, such as CRISPR/Cas9 gRNA-based genome-editing, can permanently disrupt the HIV genome. However, HIV genome-editing may accelerate viral escape, questioning the feasibility of the approach. Here, we demonstrate that CRISPR/Cas9 targeting of single HIV loci, only partially inhibits HIV replication and facilitates rapid viral escape at the target site. A combinatorial approach of two strong gRNAs targeting different regions of the HIV genome can completely abrogate viral replication and prevent viral escape. Our data shows that the accelerating effect of gene-editing on viral escape can be overcome and as such gene-editing may provide a future alternative for control of HIV-infection.
Department of Medical Microbiology, Virology, University Medical Center Utrecht, Utrecht, The Netherlands.
Robert Jan
RJ
Lebbink
Department of Medical Microbiology, Virology, University Medical Center Utrecht, Utrecht, The Netherlands.
Dorien C M
DC
de Jong
Department of Medical Microbiology, Virology, University Medical Center Utrecht, Utrecht, The Netherlands.
Femke
F
Wolters
Department of Medical Microbiology, Virology, University Medical Center Utrecht, Utrecht, The Netherlands.
Elisabeth M
EM
Kruse
Department of Medical Microbiology, Virology, University Medical Center Utrecht, Utrecht, The Netherlands.
Petra M
PM
van Ham
Department of Medical Microbiology, Virology, University Medical Center Utrecht, Utrecht, The Netherlands.
Emmanuel J H J
EJ
Wiertz
Department of Medical Microbiology, Virology, University Medical Center Utrecht, Utrecht, The Netherlands.
Monique
M
Nijhuis
D064113:CRISPR-Cas Systems; D000072669:Gene Editing / Q000379:methods*; D018390:Gene Targeting; D016679:Genome, Viral; D057809:HEK293 Cells; D015658:HIV Infections / Q000628:therapy* / Q000821:virology; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D019169:Jurkat Cells; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
D000094704:RNA, Guide, CRISPR-Cas Systems