CRISPR artificial splicing factors

Alternative splicing allows expression of mRNA isoforms from a single gene, expanding the diversity of the proteome. Its prevalence in normal biological and disease processes warrant precise tools for modulation. Here we report the engineering of CRISPR Artificial Splicing Factors (CASFx) based on R...

Descripción completa

Detalles Bibliográficos
Autores principales: Du, Menghan, Jillette, Nathaniel, Zhu, Jacqueline Jufen, Li, Sheng, Cheng, Albert Wu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7293279/
https://www.ncbi.nlm.nih.gov/pubmed/32532987
http://dx.doi.org/10.1038/s41467-020-16806-4
_version_ 1783546268891480064
author Du, Menghan
Jillette, Nathaniel
Zhu, Jacqueline Jufen
Li, Sheng
Cheng, Albert Wu
author_facet Du, Menghan
Jillette, Nathaniel
Zhu, Jacqueline Jufen
Li, Sheng
Cheng, Albert Wu
author_sort Du, Menghan
collection PubMed
description Alternative splicing allows expression of mRNA isoforms from a single gene, expanding the diversity of the proteome. Its prevalence in normal biological and disease processes warrant precise tools for modulation. Here we report the engineering of CRISPR Artificial Splicing Factors (CASFx) based on RNA-targeting CRISPR-Cas systems. We show that simultaneous exon inclusion and exclusion can be induced at distinct targets by differential positioning of CASFx. We also create inducible CASFx (iCASFx) using the FKBP-FRB chemical-inducible dimerization domain, allowing small molecule control of alternative splicing. Finally, we demonstrate the activation of SMN2 exon 7 splicing in spinal muscular atrophy (SMA) patient fibroblasts, suggesting a potential application of the CASFx system.
format Online
Article
Text
id pubmed-7293279
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-72932792020-06-16 CRISPR artificial splicing factors Du, Menghan Jillette, Nathaniel Zhu, Jacqueline Jufen Li, Sheng Cheng, Albert Wu Nat Commun Article Alternative splicing allows expression of mRNA isoforms from a single gene, expanding the diversity of the proteome. Its prevalence in normal biological and disease processes warrant precise tools for modulation. Here we report the engineering of CRISPR Artificial Splicing Factors (CASFx) based on RNA-targeting CRISPR-Cas systems. We show that simultaneous exon inclusion and exclusion can be induced at distinct targets by differential positioning of CASFx. We also create inducible CASFx (iCASFx) using the FKBP-FRB chemical-inducible dimerization domain, allowing small molecule control of alternative splicing. Finally, we demonstrate the activation of SMN2 exon 7 splicing in spinal muscular atrophy (SMA) patient fibroblasts, suggesting a potential application of the CASFx system. Nature Publishing Group UK 2020-06-12 /pmc/articles/PMC7293279/ /pubmed/32532987 http://dx.doi.org/10.1038/s41467-020-16806-4 Text en © The Author(s) 2020 Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Du, Menghan
Jillette, Nathaniel
Zhu, Jacqueline Jufen
Li, Sheng
Cheng, Albert Wu
CRISPR artificial splicing factors
title CRISPR artificial splicing factors
title_full CRISPR artificial splicing factors
title_fullStr CRISPR artificial splicing factors
title_full_unstemmed CRISPR artificial splicing factors
title_short CRISPR artificial splicing factors
title_sort crispr artificial splicing factors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7293279/
https://www.ncbi.nlm.nih.gov/pubmed/32532987
http://dx.doi.org/10.1038/s41467-020-16806-4
work_keys_str_mv AT dumenghan crisprartificialsplicingfactors
AT jillettenathaniel crisprartificialsplicingfactors
AT zhujacquelinejufen crisprartificialsplicingfactors
AT lisheng crisprartificialsplicingfactors
AT chengalbertwu crisprartificialsplicingfactors

Ejemplares similares