1. Protoplasts: From Isolation to CRISPR/Cas Genome Editing Application
- Author
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Jin-Jun Yue, Jin-Ling Yuan, Fu-Hui Wu, Yu-Hsuan Yuan, Qiao-Wei Cheng, Chen-Tran Hsu, and Choun-Sea Lin
- Subjects
RNP ,transient transfection ,DNA-free ,CRISPR/cas (clustered regularly interspaced short palindromic repeats) ,protoplasts ,Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
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.
- Published
- 2021
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