Protoplasts: From Isolation to CRISPR/Cas Genome Editing Application

In the clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas) system, protoplasts are not only useful for rapidly validating the mutagenesis efficiency of various RNA-guided endonucleases, promoters, sgRNA designs, or Cas proteins, but can also be a platf...

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Bibliographic Details
Published in:Frontiers in genome editing 2021-08, Vol.3, p.717017
Main Authors: Yue, Jin-Jun, Yuan, Jin-Ling, Wu, Fu-Hui, Yuan, Yu-Hsuan, Cheng, Qiao-Wei, Hsu, Chen-Tran, Lin, Choun-Sea
Format: Article
Language:English
Subjects:
CRISPR/cas (clustered regularly interspaced short palindromic repeats)
DNA-free
Genome Editing
protoplasts
RNP
transient transfection
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Summary:In the clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas) system, protoplasts are not only useful for rapidly validating the mutagenesis efficiency of various RNA-guided endonucleases, promoters, sgRNA designs, or Cas proteins, but can also be a platform for DNA-free gene editing. To date, the latter approach has been applied to numerous crops, particularly those with complex genomes, a long juvenile period, a tendency for heterosis, and/or self-incompatibility. Protoplast regeneration is thus a key step in DNA-free gene editing. In this report, we review the history and some future prospects for protoplast technology, including protoplast transfection, transformation, fusion, regeneration, and current protoplast applications in CRISPR/Cas-based breeding.
ISSN:2673-3439
2673-3439
DOI:10.3389/fgeed.2021.717017