Cas-CLOVER Targeted Editing
The dimeric targeted nuclease technology, Cas-CLOVER, differs from CRISPR/Cas9 as it has limited off-target effects and clear commercial freedom-to-operate. Cas-CLOVER was validated with high activity in plants by knocking out a commercially valuable gene in tobacco. We are focused on partnering with groups who can apply this technology broadly in plant trait engineering.
Precision Genome Editing in Rice
As is most plants, homologous recombination (HR) mediated precision genome editing in rice is an infrequent event. Efficiency can be vastly improved by positive-negative selection. However, the positive selection marker (HPT) needs to be seamlessly removed from the edited locus. PiggyBac is the ideal technology for this task as it can be used to introduce the desired gene edit, followed by footprint-free excision after selection.
Herbicide Resistance Example
piggyBac based gene editing was achieved at the rice ALS locus. The targeting vector carries selection markers flanked by piggyBac target sequences and the desired mutations are located in the arms of homology. Following selection of the edit and footprint-free excision, the ALS gene contains the desired point mutations with all other components of the targeting seamlessly removed. The result is rice with point mutations are resistant to the herbicide bispyribac sodium (BS).