CRISPR-GRIT: Guide RNAs with Integrated Repair Templates Enable Precise Multiplexed Genome Editing in the Diploid Fungal Pathogen Candida albicans

Cotter, Christopher J.; Trinh, Cong T.

doi:10.1089/crispr.2024.0052

CRISPR-GRIT: Guide RNAs with Integrated Repair Templates Enable Precise Multiplexed Genome Editing in the Diploid Fungal Pathogen Candida albicans

Journal Article · Tue Oct 22 00:00:00 EDT 2024 · The CRISPR Journal

DOI:https://doi.org/10.1089/crispr.2024.0052· OSTI ID:2479302

Cotter, Christopher J. ^[1]; ^[1]

University of Tennessee, Knoxville, TN (United States)

Candida albicans, an opportunistic fungal pathogen, causes severe infections in immunocompromised individuals. Limited classes and overuse of current antifungals have led to the rapid emergence of antifungal resistance. Thus, there is an urgent need to understand fungal pathogen genetics to develop new antifungal strategies. Genetic manipulation of C. albicans is encumbered by its diploid chromosomes requiring editing both alleles to elucidate gene function. Although the recent development of CRISPR-Cas systems has facilitated genome editing in C. albicans, large-scale and multiplexed functional genomic studies are still hindered by the necessity of cotransforming repair templates for homozygous knockouts. Here, we present CRISPR-GRIT (Guide RNAs with Integrated Repair Templates), a repair template-integrated guide RNA design for expedited gene knockouts and multiplexed gene editing in C. albicans. Here, we envision that this method can be used for high-throughput library screens and identification of synthetic lethal pairs in both C. albicans and other diploid organisms with strong homologous recombination machinery.

View Accepted Manuscript (DOE)

Research Organization:: University of Tennessee, Knoxville, TN (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0019412

OSTI ID:: 2479302

Journal Information:: The CRISPR Journal, Journal Name: The CRISPR Journal Journal Issue: 6 Vol. 7; ISSN 2573-1599

Publisher:: Mary Ann Liebert, Inc.Copyright Statement

Country of Publication:: United States

Language:: English

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Related Subjects

60 APPLIED LIFE SCIENCES
CRISPR-Cas
CRISPR-GRIT
Candida albicans
diploid
genetic interaction
homologous recombination
library screen
multiplexing
multiploidy
repair templates

CRISPR-GRIT: Guide RNAs with Integrated Repair Templates Enable Precise Multiplexed Genome Editing in the Diploid Fungal Pathogen Candida albicans

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References (31)

Figures / Tables (11)

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