Genome-wide Characterization of Diverse Bacteriophages Enabled by RNA-Binding CRISPRi
Genome-wide Characterization of Diverse Bacteriophages Enabled by RNA-Binding CRISPRi
Bacteriophages constitute one of the largest sources of unknown gene content in the biosphere. Even for well-studied model phages, robust experimental approaches to identify and study their essential genes remain elusive. We uncover and exploit the conserved vulnerability of the phage transcriptome to facilitate genome-wide protein expression knockdown via programmable RNA-binding protein dRfxCas13d (CRISPRi-ART) across diverse phages and their host. Establishing the first broad-spectrum phage functional genomics platform, we predict over 90 essential genes across four phage genomes, a third of which have no known function. These results highlight hidden infection strategies encoded in the most abundant biological entities on earth and provide a facile platform to study them.
Schoeniger Joseph S Dr.、Patel Jaymin R Dr.、Colognori David Dr.、Lahiri Arushi、Trinidad Marena、Boger Ron、Beurnier Sebastien、Doudna Jennifer A Dr.、Adler Benjamin A Dr.、Savage David F Dr.、Cress Brady F Dr.、Charles Emeric J Dr.、Cui Michael L、Al-Shimary Muntathar J、Miller Kate V、Ambruster Emily、Barrangou Rodolphe Dr.、Nomburg Jason Dr.、Mutalik Vivek K Dr.、Pogliano Joseph A Dr.
生物科学研究方法、生物科学研究技术遗传学分子生物学
Schoeniger Joseph S Dr.,Patel Jaymin R Dr.,Colognori David Dr.,Lahiri Arushi,Trinidad Marena,Boger Ron,Beurnier Sebastien,Doudna Jennifer A Dr.,Adler Benjamin A Dr.,Savage David F Dr.,Cress Brady F Dr.,Charles Emeric J Dr.,Cui Michael L,Al-Shimary Muntathar J,Miller Kate V,Ambruster Emily,Barrangou Rodolphe Dr.,Nomburg Jason Dr.,Mutalik Vivek K Dr.,Pogliano Joseph A Dr..Genome-wide Characterization of Diverse Bacteriophages Enabled by RNA-Binding CRISPRi[EB/OL].(2025-03-28)[2025-07-16].https://www.biorxiv.org/content/10.1101/2023.09.18.558157.点此复制
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