Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems

CRISPR/Cas-mediated genome editing in human pluripotent stem cells (hPSCs) offers unprecedented opportunities for developing in vitro disease modeling, drug screening and cell-based therapies. To efficiently deliver the CRISPR components, here we developed two all-in-one vectors containing Cas9/gRNA...

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Autores principales: Jiang, Yuqian, Hoenisch, Rachel Catherine, Chang, Yun, Bao, Xiaoping, Cameron, Craig E., Lian, Xiaojun Lance
Formato: Online Artículo Texto
Lenguaje:English
Publicado: KeAi Publishing 2022
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8964983/
https://www.ncbi.nlm.nih.gov/pubmed/35386818
http://dx.doi.org/10.1016/j.bioactmat.2022.01.046
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author Jiang, Yuqian
Hoenisch, Rachel Catherine
Chang, Yun
Bao, Xiaoping
Cameron, Craig E.
Lian, Xiaojun Lance
author_facet Jiang, Yuqian
Hoenisch, Rachel Catherine
Chang, Yun
Bao, Xiaoping
Cameron, Craig E.
Lian, Xiaojun Lance
author_sort Jiang, Yuqian
collection PubMed
description CRISPR/Cas-mediated genome editing in human pluripotent stem cells (hPSCs) offers unprecedented opportunities for developing in vitro disease modeling, drug screening and cell-based therapies. To efficiently deliver the CRISPR components, here we developed two all-in-one vectors containing Cas9/gRNA and inducible Cas13d/gRNA cassettes for robust genome editing and RNA interference respectively. These vectors utilized the PiggyBac transposon system, which allows stable expression of CRISPR components in hPSCs. The Cas9 vector PB-CRISPR exhibited high efficiency (up to 99%) of inducing gene knockout in both protein-coding genes and long non-coding RNAs. The other inducible Cas13d vector achieved extremely high efficiency in RNA knockdown (98% knockdown for CD90) with optimized gRNA designs. Taken together, our PiggyBac CRISPR vectors can serve as powerful toolkits for studying gene functions in hPSCs.
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spelling pubmed-89649832022-04-05 Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems Jiang, Yuqian Hoenisch, Rachel Catherine Chang, Yun Bao, Xiaoping Cameron, Craig E. Lian, Xiaojun Lance Bioact Mater Article CRISPR/Cas-mediated genome editing in human pluripotent stem cells (hPSCs) offers unprecedented opportunities for developing in vitro disease modeling, drug screening and cell-based therapies. To efficiently deliver the CRISPR components, here we developed two all-in-one vectors containing Cas9/gRNA and inducible Cas13d/gRNA cassettes for robust genome editing and RNA interference respectively. These vectors utilized the PiggyBac transposon system, which allows stable expression of CRISPR components in hPSCs. The Cas9 vector PB-CRISPR exhibited high efficiency (up to 99%) of inducing gene knockout in both protein-coding genes and long non-coding RNAs. The other inducible Cas13d vector achieved extremely high efficiency in RNA knockdown (98% knockdown for CD90) with optimized gRNA designs. Taken together, our PiggyBac CRISPR vectors can serve as powerful toolkits for studying gene functions in hPSCs. KeAi Publishing 2022-02-07 /pmc/articles/PMC8964983/ /pubmed/35386818 http://dx.doi.org/10.1016/j.bioactmat.2022.01.046 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Jiang, Yuqian
Hoenisch, Rachel Catherine
Chang, Yun
Bao, Xiaoping
Cameron, Craig E.
Lian, Xiaojun Lance
Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems
title Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems
title_full Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems
title_fullStr Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems
title_full_unstemmed Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems
title_short Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems
title_sort robust genome and rna editing via crispr nucleases in piggybac systems
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8964983/
https://www.ncbi.nlm.nih.gov/pubmed/35386818
http://dx.doi.org/10.1016/j.bioactmat.2022.01.046
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