Genome editing with FnCas12a using crRNA with altered target length in Oryza sativa. Oryza sativa Japonica Group

The crRNA of FnCas12a generally has a 24-nt guide sequence that is complementary to the target DNA sequence. Previous reports and our studies have shown that changing the length of guide sequence affects genome editing efficiency in rice. The goal of this project is to reveal the effects of guide sequence length on mutation pattern and efficiency. We constructed crRNAs with 18-nt, 24-nt, and 30-nt guide sequence and assessed the deletion mutation pattern and mutation efficiency at five target sites in rice genes by amplicon deep sequencing analysis. This study will facilitate establishing a high efficiently and precise CRISPR/FnCas12a-mediated genome editing system.