CRISPR-Cas9-mediated Mutagenesis in Physcomitrella patens
November 9, 2016| |
The ability to address the CRISPR-Cas9 nuclease complex to any target DNA using customizable guide RNAs has allowed genome engineering in several species. Scientists from the INRA Centre de Versailles-Grignon now describe the first successful use of CRISPR-Cas9 in a nonvascular plant, the moss Physcomitrella patens.
The team designed single-guide RNAs (sgRNAs) to target an endogenous reporter gene, PpAPT, which, when inactive, confers resistance to 2-fluoroadenine. Transformation of moss protoplasts with these sgRNAs and the Cas9 coding sequence from Streptococcus pyogenes triggered mutagenesis at the PpAPT gene in about 2% of the regenerated plants. The resulting transformants exhibited resistance to 2-fluoroadenine.
Deletions were mainly observed, with most of them resulting from alternative end-joining-driven repairs. For more on this promising study, read the article in Plant Biotechnology Journal.
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