A new and novel high-fidelity genome editing tool for banana using Cas-CLOVER

Genome editing with engineered nucleases has developed rapidly in the last decade and has transformed researchers' ability to manipulate various organisms' genomes. Among the gene editing technologies, CRISPR/Cas9 has been adopted as the most popular technique for functional genomics and crop improvement due to its ease of use, design flexibility, high efficiency, and multiplexing. However, the intellectual property (IP) surrounding this technology is complex and is difficult to license for growers. Demeetra’s alternative to CRISPR/Cas9 is Cas-CLOVER. In this communication, Leena Tripathi’s lab establishes Cas-CLOVER as an alternative genome-editing tool to CRISPR/Cas9 by targeting MusaPDS in the banana genome. Albino banana shoots were regenerated from embryogenic cell suspensions (ECS) and the desired mutations within the target site were sequence verified. In addition to regenerating completely albino plants, Cas-CLOVER creates larger deletions than CRISPR/Cas9. The Cas-CLOVER system is an alternative genome-editing tool that has the efficiency of CRISPR/Cas9 and improved fidelity but due to its clear IP and simple license, has the potential to facilitate the development of genome-edited crops quickly and with full commercial freedom.

Fig 1d 4-6

Figure 1d 4-6:

Regeneration genome edited banana events using the Cas-CLOVER technology targeting the MusaPDS gene. 4) Embryo germination for transformed ECS; 5) Control non-edited shoots; 6) Albino edited events.