Lambert Lab CRISPR-Cas12a Xseq Seed Region Crosstalk Catalytically Active
A compact synthetic Cas12a inverter containing randomized bases is used to assay the cutting efficacy of a DNA target. All components are contained  within a plasmid, and the experiment is conducted in E. coli. Here, the target is a DNA sequence containing a 6bp randomized region, which  corresponds to the Cas12a first 6 bp 'seed' region. The crRNA also has a 6bp randomized region in the corresponding location. Cas12a cutting efficacy is  measured via growth using plasmids with (ActiveCas) and lacking (Control) catalytically-active Cas12a due to the fitness disadvantage of cleavage of  the plasmid containing antibiotic resistance (Kanamycin).
SUB5844072
ActiveCas