Arrayed CRISPR gRNA Libraries

Screen Smarter Not Harder. Effective Knockout Strategy for Better CRISPR Screening.

Other loss-of-function CRISPR screening libraries are often plagued with laborious preparation, data analysis, poor editing efficiency and low quality reagents that can be cytotoxic to more sensitive cell types such as primary cells. Our Arrayed CRISPR gRNA library options are different.

Powered by EditCo’s unique multi-guide design, our Arrayed CRISPR gRNA Libraries are comprised of SpCas9 sgRNAs targeting one gene per well in a multiwell plate format. The multi-guide design consists of up to three spatially coordinated SpCas9 sgRNAs that induce fragment deletions in an early single exon of a gene. Large fragment deletions in the target gene increased likelihood of functional knockout by not relying on random indel creation. Overall, this decreases the likelihood of false negatives, reliably knocking out any human or mouse protein-coding gene. 

Our CRISPR screening libraries are ready-to-transfect, either complexed into RNPs or directly into a Cas9-expressing cell line avoiding long preparation protocols. The arrayed format avoids the need of messy next-generation sequencing (NGS) data deconvolution to interpret your screening results and allows to perform binary and multiparametric functional assays. Furthermore, you can analyze your edits in seconds using Sanger sequencing with our CE tool. Our unique multi-guide design ensures you can genotype with one single reaction.

Figure 1. EditCo Arrayed gRNA Library Workflow. Incorporating smart guide design with automation coupled with our high-quality control, we plate only the best-quality sgRNA into each well with high precision enabling you to reliably knock out your targets in your screens with confidence, we guarantee it (excluding non-essential genes). Due to our optimized workflow, we can deliver your Arrayed CRISPR gRNA Libraries in as few as 7 days at unrivaled speed.

Available Gene Pathways

How Does EditCo's unique guide design induce a fragment deletion?

EditCo’s multi-guide design is composed of up to 3 sgRNAs targeting an early single exon of a gene of interest. The guides are spatially coordinated to induce a guided repair that results in fragment deletions. The guide spacing was also designed to guarantee effective analysis of your CRISPR edits with one single reaction and in seconds using Sanger sequencing with our free unique bioinformatics tool, Inference of CRISPR Edits (ICE).

Consistent and Reliable Knockouts

Figure 3. Better knockout efficiency was found across 32 genetic targets assessed. EditCo’s multi-guide design consistently results in high Knockout (KO) Scores across 32 genetic targets assessed compared to strategies that rely on individual gRNAs. gRNAs displayed 29.2% better median knockout efficiency when introduced in a multi-guide format (89.9% KO score) relative to an individual sgRNA format (69.6% KO Score).

Achieve Robust Genotype and Phenotype Screening Results

Figure 2. Across 77 genes, arrayed multi-guide library resulted in an average Knockout Score of 75%. Using our multi-guide algorithm, up to 3 modified sgRNA per target (across 86 genes) were transfected into U2OS-Cas9 expressing cell line using nucleofection. This resulted in an average Knockout Score of 75% for 77 of the genes. Nine of the 86 genes failed due to sequencing errors.

Figure 3. Western blot analyses for all 5 knockout target genes indicate a complete depletion of proteins across the 3 specified time points (days 7, 14, and 21) relative to the negative controls. Knockout cell pools for the target genes were generated by nucleofecting U2OS cells with multi-guide sgRNA and Cas9 (as RNPs). A negative control pool was also transfected for each target using non-targeting sgRNA.

EditCo Libraries Enables High Editing Efficiencies in a Large-scale Primary Cell Screen

Figure 4. Screening showed a 97% median editing efficiency using across 280 loci. Multi-guide sgRNA was transfected into primary dendritic cells using nucleofection. Editing efficiency was assessed via NGS across 280 loci. Data courtesy of Weissman Lab, UCSF.