This technology utilizes reprogrammed bacterial cells to efficiently and cost-effectively generate highly comprehensive, genome-wide crRNA libraries for CRISPR screens. crRNA is the canonical form of guide RNA (gRNA) that has been engineered and widely used.
The current method of generating gRNA libraries for genome-wide CRISPR screens is pooled oligonucleotide synthesis. This method can be costly, labor-intensive and time-consuming, with a turnaround of 1-2 weeks or longer. Furthermore, this method generates libraries with low diversity, substantially limiting the coverage and sensitivity of CRISPR screens.
This technology reprograms the S. pyogenes CRISPR-Cas system to convert bacterial cells into “factories” that generate hundreds of thousands of crRNAs in as short as a single day. The resulting crRNA library covers up to 95% of the targeted microbial genome, with the average gene targeted by more than 100 distinct crRNAs. This method can generate both single and dual crRNA libraries to elucidate genetic interactions and record the evolutionary trajectory of acquired traits.
This technology has been validated with S. aureus, E. coli, and wild-type bacterial pathogens.
Patent Pending
IR CU19311
Licensing Contact: Joan Martinez