Journal Article

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

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

2020

Overview

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.

• Publisher: BioMed Central,Springer Nature B.V,BMC
• Subject: Adaptation / Adenosine Deaminase - metabolism / Animal Genetics and Genomics / Animals / Bioinformatics / Biomedical and Life Sciences / Cas13 / CasRX / Cell culture / CRISPR / CRISPR-Associated Proteins - metabolism / CRISPR-Cas Systems / crRNA design / Drosophila / Drosophila - genetics / Efficiency / Endonuclease / Endonucleases - metabolism / Enzymes / Evolutionary Biology / Gene Expression / Genes / Genetic engineering / genome / Genomes / Human Genetics / Insects / Kinases / Life Sciences / Method / Microbial Genetics and Genomics / Mitochondria / Plant Genetics and Genomics / Post-transcription / Proteins / Ribonucleic acid / RNA / RNA - genetics / RNA manipulation / RNA Processing, Post-Transcriptional / RNA, Mitochondrial / RNA-Binding Proteins - metabolism