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Anti‐HIV‐1 potency of the CRISPR/Cas9 system insufficient to fully inhibit viral replication
Author(s) -
Ueda Shuhei,
Ebina Hirotaka,
Kanemura Yuka,
Misawa Naoko,
Koyanagi Yoshio
Publication year - 2016
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/1348-0421.12395
Subject(s) - crispr , guide rna , biology , cas9 , genome editing , virology , viral replication , viral vector , lentivirus , genome , computational biology , rna , vector (molecular biology) , human immunodeficiency virus (hiv) , genetics , virus , gene , recombinant dna , viral disease
The range of genome‐editing tools has recently been expanded. In particular, an RNA‐guided genome‐editing tool, the clustered regularly interspaced short palindromic repeat (CRISPR)‐associated 9 (Cas9) system, has many applications for human diseases. In this study, guide RNA (gRNA) to target gag , pol and a long terminal repeat of HIV‐1 was designed and used to generate gRNA‐expressing lentiviral vectors. An HIV‐1‐specific gRNA and Cas9 were stably dually transduced into a highly HIV‐1‐susceptible human T‐cell line and the inhibitory ability of the anti‐HIV‐1 CRISPR/Cas9 lentiviral vector assessed. Although clear inhibition of the early phase of HIV‐1 infection was observed, as evaluated by a VSV‐G‐pseudotyped HIV‐1 reporter system, the anti‐HIV‐1 potency in multiple rounds of wild type (WT) viral replication was insufficient, either because of generation of resistant viruses or overcoming of the activity of the WT virus. Thus, there are potential difficulties that must be addressed when considering anti‐HIV‐1 treatment with the CRISPR/Cas9 system alone.