期刊
FRONTIERS IN GENOME EDITING
卷 3, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fgeed.2021.717017
关键词
RNP; transient transfection; DNA-free; CRISPR; cas (clustered regularly interspaced short palindromic repeats); protoplasts
Protoplasts play a crucial role in validating gene editing tools and conducting DNA-free gene editing in crops with complex genomes, long juvenile periods, heterosis, and/or self-incompatibility in the CRISPR/Cas system. The applications of protoplast technology in CRISPR/Cas-based breeding have significant implications for crop improvement.
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.
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