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Regulation of Cas9 by viral proteins Tat and Rev for HIV-1 inactivation
- Source :
- Antiviral Research. 180:104856
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- While combined antiretroviral therapy (cART) has had a great impact on the treatment of HIV-1 infection, the persistence of long-lived cells with an intact provirus precludes virus eradication and sterilizing cure. CRISPR/Cas9 genome editing has become an efficient tool to eradicate HIV-1 genome or prevent replication. Furthermore, regulation of Cas9 gene expression by HIV can induce mutations that could inactivate the proviral genome, making a gene therapy safe by preventing the induction of non-specific mutations, which could compromise the integrity of healthy cells. In this study, isolated HIV-1 LTR, INS and RRE sequences were used to regulate Cas9 expression in HEK293 cells, and guide RNAs (gRNAs) were designed to target mutations in HIV-1 conserved regions such as tat and rev regulatory genes. We demonstrate that Cas9 expression in our system is controlled by the HIV-1 Tat and Rev proteins, leading to self-regulation of gene edition, and showing a strong antiviral effect by inactivating HIV-1 replication. Sequencing analysis confirmed that viral genome was partially excised by multiplex editing (90% efficiency), and viral capsid protein (CA-p24) was undetectable. In conclusion, the self-regulated CRISPR/Cas9 system may be a reliable and accurate strategy for eliminating HIV-1 infection whose effect will be restricted to infected cells.
- Subjects :
- Gene Expression Regulation, Viral
0301 basic medicine
viruses
030106 microbiology
Biology
Virus Replication
Genome
Virus
03 medical and health sciences
Genome editing
CRISPR-Associated Protein 9
Virology
Humans
CRISPR
Gene
Regulator gene
Gene Editing
Pharmacology
Cas9
rev Gene Products, Human Immunodeficiency Virus
Provirus
HEK293 Cells
030104 developmental biology
HIV-1
Virus Inactivation
tat Gene Products, Human Immunodeficiency Virus
CRISPR-Cas Systems
RNA, Guide, Kinetoplastida
Subjects
Details
- ISSN :
- 01663542
- Volume :
- 180
- Database :
- OpenAIRE
- Journal :
- Antiviral Research
- Accession number :
- edsair.doi.dedup.....074fb49a373244505acbabfa4c3528d2