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Open AccessArticle

CRISPR-Cas9 Dual-gRNA Attack Causes Mutation, Excision and Inversion of the HIV-1 Proviral DNA

Laboratory of Experimental Virology, Department of Medical Microbiology, Amsterdam UMC, University of Amsterdam, 1105AZ Amsterdam, The Netherlands
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Authors to whom correspondence should be addressed.
Viruses 2020, 12(3), 330; https://doi.org/10.3390/v12030330
Received: 20 February 2020 / Revised: 11 March 2020 / Accepted: 13 March 2020 / Published: 18 March 2020
(This article belongs to the Special Issue Antiretroviral Drug Development and HIV Cure Research)
Although several studies demonstrated that the HIV proviral DNA can be effectively targeted and inactivated by the CRISPR-Cas9 system, the precise inactivation mechanism has not yet been analyzed. Whereas some studies suggested efficient proviral DNA excision upon dual-gRNA/Cas9 treatment, we previously demonstrated that hypermutation of the target sites correlated with permanent virus inactivation. To better understand the mechanism underlying HIV inactivation, we analyzed the proviral DNA upon Cas9 attack with gRNA pairs. We observed that dual-gRNA targeting resulted more frequently in target site mutation than fragment excision, while fragment inversion was rarely observed. The frequencies varied for different gRNA combinations without an obvious relationship with the distance between the target sites, indicating that other gRNA and target DNA characteristics influence the DNA cleavage and repair processes. View Full-Text
Keywords: HIV; CRISPR-Cas; excision; mutation; DNA repair HIV; CRISPR-Cas; excision; mutation; DNA repair
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MDPI and ACS Style

Binda, C.S.; Klaver, B.; Berkhout, B.; Das, A.T. CRISPR-Cas9 Dual-gRNA Attack Causes Mutation, Excision and Inversion of the HIV-1 Proviral DNA. Viruses 2020, 12, 330. https://doi.org/10.3390/v12030330

AMA Style

Binda CS, Klaver B, Berkhout B, Das AT. CRISPR-Cas9 Dual-gRNA Attack Causes Mutation, Excision and Inversion of the HIV-1 Proviral DNA. Viruses. 2020; 12(3):330. https://doi.org/10.3390/v12030330

Chicago/Turabian Style

Binda, Caroline S.; Klaver, Bep; Berkhout, Ben; Das, Atze T. 2020. "CRISPR-Cas9 Dual-gRNA Attack Causes Mutation, Excision and Inversion of the HIV-1 Proviral DNA" Viruses 12, no. 3: 330. https://doi.org/10.3390/v12030330

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