Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems

Yuqian Jiang; Rachel Catherine Hoenisch; Yun Chang; Xiaoping Bao; Craig E. Cameron; Xiaojun Lance Lian

doi:10.1016/j.bioactmat.2022.01.046

Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems

Yuqian Jiang, Rachel Catherine Hoenisch, Yun Chang, Xiaoping Bao, Craig E. Cameron, Xiaojun Lance Lian

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	313-320
Number of pages	8
Journal	Bioactive Materials
Volume	14
DOIs	https://doi.org/10.1016/j.bioactmat.2022.01.046
State	Published - Aug 2022

All Science Journal Classification (ASJC) codes

Biotechnology
Biomaterials
Biomedical Engineering

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.bioactmat.2022.01.046

Cite this

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title = "Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems",

abstract = "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.",

author = "Yuqian Jiang and Hoenisch, {Rachel Catherine} and Yun Chang and Xiaoping Bao and Cameron, {Craig E.} and Lian, {Xiaojun Lance}",

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year = "2022",

month = aug,

doi = "10.1016/j.bioactmat.2022.01.046",

language = "English (US)",

volume = "14",

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AU - Jiang, Yuqian

AU - Hoenisch, Rachel Catherine

AU - Chang, Yun

AU - Bao, Xiaoping

AU - Cameron, Craig E.

AU - Lian, Xiaojun Lance

PY - 2022/8

Y1 - 2022/8

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AB - 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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Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems

Abstract

All Science Journal Classification (ASJC) codes

UN SDGs

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