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

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 Ca...

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Detalles Bibliográficos
Autores principales: Huynh, Nhan, Depner, Noah, Larson, Raegan, King-Jones, Kirst
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7670108/
https://www.ncbi.nlm.nih.gov/pubmed/33203452
http://dx.doi.org/10.1186/s13059-020-02193-y
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author Huynh, Nhan
Depner, Noah
Larson, Raegan
King-Jones, Kirst
author_facet Huynh, Nhan
Depner, Noah
Larson, Raegan
King-Jones, Kirst
author_sort Huynh, Nhan
collection PubMed
description 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-020-02193-y.
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spelling pubmed-76701082020-11-17 A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila Huynh, Nhan Depner, Noah Larson, Raegan King-Jones, Kirst Genome Biol Method 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-020-02193-y. BioMed Central 2020-11-17 /pmc/articles/PMC7670108/ /pubmed/33203452 http://dx.doi.org/10.1186/s13059-020-02193-y Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Method
Huynh, Nhan
Depner, Noah
Larson, Raegan
King-Jones, Kirst
A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila
title A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila
title_full A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila
title_fullStr A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila
title_full_unstemmed A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila
title_short A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila
title_sort versatile toolkit for crispr-cas13-based rna manipulation in drosophila
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7670108/
https://www.ncbi.nlm.nih.gov/pubmed/33203452
http://dx.doi.org/10.1186/s13059-020-02193-y
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