Protoplasts: From Isolation to CRISPR/Cas Genome Editing Application

• Published in: Frontiers in genome editing 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: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 11.08.2021
• Subjects: CRISPR/cas (clustered regularly interspaced short palindromic repeats); DNA-free; Genome Editing; protoplasts; RNP; transient transfection; transient transfection; CRISPR/cas (clustered regularly interspaced short palindromic repeats); DNA-free; protoplasts; RNP
• ISSN: 2673-3439; 2673-3439
• DOI: 10.3389/fgeed.2021.717017

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.