Improvement of genome editing efficiency by Cas9 codon optimization in Japanese cedar (<i>Cryptomeria japonica</i> D. Don).

ABSTRACT: Japanese cedar or sugi (Cryptomeria japonica D. Don) is among the most important plantation conifers in Japan, occupying 12% of the total land area in the country. We have successfully established a CRISPR/Cas9-based genome editing system in C. japonica. However, in practical use, we encountered problems of low efficiency when generating biallelic mutations, i.e., target gene knockouts. As part of our efforts to improve efficiency, we codon-optimized the Cas9 gene, evaluated by the genome editing efficiency of CjChl I, a gene encoding a chlorophyll biosynthesis enzyme. As a result, our codon-optimized SpCas9, named ^CjSpCas9, performed the highest genome editing efficiency of two targets (t4, t1+t2). Specifically, the biallelic disruption efficiency of the

SUBMITTER: Nanasato Y 

PROVIDER: S-EPMC11897717 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature