FLI-Seq

CRISPR screens are large-scale loss-of-function screens that are used to identify key genes responsible for a particular function or phenotype of a cell. After CRISPR screening, genomic DNA is harvested from the selected pool of cells and PCR is performed to isolate and identify the single guide RNA (sgRNA) sequences resulting from the screen. Fast Library Insert Sequencing, or FLI-Seq™, facilitates the efficient generation of sgRNA libraries resulting from CRISPR screens by enriching for the relevant sgRNA-encoding gDNA regions. The FLI-Seq method generates libraries with far fewer PCR reactions and PCR cycles, thereby reducing PCR duplication and PCR artifacts. Optimized PCR amplification primers yield library fragments beginning at sgRNA sequence, requiring very short read lengths and simplifying data analysis. FLI-Seq simply generates high quality sgRNA libraries with highly technical reproducibility that requires less time, less starting material, and less reagents than other approaches for processing CRISPR screens.