Improving CRISPR-Cas-mediated RNA targeting and gene editing using SPLCV replicon-based expression vectors in Nicotiana benthamiana.

In this study, we developed Sweet Potato Leaf Curl Virus (SPLCV) replicon-based expression vectors and tested the efficiency of these vectors in CRISPR-Cas-mediated RNA targeting and gene editing by using Nicotiana benthamiana as model plant. Our results showed that the SPLCV-based vectors remarkably enhance the LwaCas13a-mediated RNA targeting activity nd SpCas9/LbCas12a-mediated gene editing efficiency in Nicotiana benthamiana. Our work expands the existing list of geminiviral replicon-based vectors in plants. The SPLCV-based vectors offer an opportunity to enhance the efficiency of CRISPR-Cas-mediated gene knockdown, knockout, and knockin in plants, especially for its host sweetpotato.