CRISPR in Nannochloropsis
CRISPR/Cas9 techniques in Nannochloropsis
Production and screening of Cas9
Production and screening of sgRNA
An effective sgRNA should not have terminal modifications and needs to present in the nucleus. In Nannochloropsis sgRNAs have been produced using protein coding promoters and exogenously produced and transformed into the cells. To generate more efficient sgRNAs from RNA polymerase II promoters, self-cleaving ribozymes can be appended to the sgRNA gene.