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Assessment of anti-HIV-1 guide RNA efficacy in cells containing the viral target sequence, corresponding gRNA, and CRISPR/Cas9
Journal article   Open access   Peer reviewed

Assessment of anti-HIV-1 guide RNA efficacy in cells containing the viral target sequence, corresponding gRNA, and CRISPR/Cas9

Alexander G. Allen, Cheng-Han Chung, Stephen D. Worrell, Glad Nwaozo, Rebekah Madrid, Anthony R. Mele, Will Dampier, Michael R. Nonnemacher and Brian Wigdahl
Frontiers in genome editing, v 5
01 Apr 2023
DOI: https://doi.org/10.3389/fgeed.2023.1101483
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://doi.org/10.3389/fgeed.2023.1101483View
Published, Version of Record (VoR)CC BY V4.0,  Open

Abstract

CRISPR/Cas9 cure strategy guide RNAs HIV-1 reservoir Gene Therapy
The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 gene editing system has been shown to be effective at inhibiting human immunodeficiency virus type 1 (HIV-1). Studies have not consistently used a trackable dual reporter system to determine what cells received the Cas9/gRNA to determine the overall knockdown of HIV. Some studies have used stably transduced cells under drug selection to accomplish this goal. Here a two-color system was used that allows tracking of viral protein expression and which cells received the CRISPR/Cas9 system. These experiments ensured that each gRNA used was a perfect match to the intended target to remove this variable. The data showed that gRNAs targeting the transactivation response element (TAR) region or other highly conserved regions of the HIV-1 genome were effective at stopping viral gene expression, with multiple assays demonstrating greater than 95 percent reduction. Conversely, gRNAs targeting conserved sites of the 5’ portion of the U3 region were largely ineffective, demonstrating that the location of edits in the long terminal repeat (LTR) matter with respect to function. In addition, it was observed that a gRNA targeting Tat was effective in a T-cell model of HIV-1 latency. Taken together, these studies demonstrated gRNAs designed to highly conserved functional regions have near 100% efficacy in vitro in cells known to have received the Cas9/gRNA pair.

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13 Record Views
8 citations in Web of Science
6 citations in Scopus

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UN Sustainable Development Goals (SDGs)

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#3 Good Health and Well-Being

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Collaboration types
Domestic collaboration
Web of Science research areas
Biotechnology & Applied Microbiology
Genetics & Heredity
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