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A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila

Nhan Huynh, Noah Depner, Raegan Larson, Kirst King‐Jones

2020Genome biology124 citationsDOIOpen Access PDF

Abstract

Advances in CRISPR technology have immensely improved our ability to manipulate nucleic acids, and the recent discovery of the RNA-targeting endonuclease Cas13 adds even further functionality. Here, we show that Cas13 works efficiently in Drosophila, both ex vivo and in vivo. We test 44 different Cas13 variants to identify enzymes with the best overall performance and show that Cas13 could target endogenous Drosophila transcripts in vivo with high efficiency and specificity. We also develop Cas13 applications to edit mRNAs and target mitochondrial transcripts. Our vector collection represents a versatile tool collection to manipulate gene expression at the post-transcriptional level.

Topics & Concepts

BiologyCRISPRComputational biologyRNA editingGeneEndonucleaseRNAmicroRNAGeneticsCRISPR and Genetic EngineeringRNA and protein synthesis mechanismsAdvanced biosensing and bioanalysis techniques
A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila | Litcius