The Dual-sgRNA Libraries for CRISPRa and CRISPRi Genetic Screens will provide augmented tools for researchers and scientists in the genome editing sector.
FREMONT, CA: The advancements in genome editing technologies have enabled scientists and researchers to add, remove, and alter specific locations of the genome with more ease. The potential promises of cost-effectiveness and speed offered by the latest technology CRISPR-Cas9 in genome editing caused a lot of excitement in the scientific community.
To augment the functionality of the CRISPR technology, Cellecta Inc. recently launched the first commercially available dual-sgRNA libraries. It is designed for CRISPR activation (CRISPRa) and CRISPR interference/repression of genes to develop enhanced results from genetic screens.
CRISPR activation (CRISPRa) and CRISPR inhibition (CRISPRi) systems are the modified versions of the CRISPR-Cas9 system. They leverage the deactivated Cas9 (dCas9) protein, which allows transcription repressor factors to be targeted to the promoter region of the gene, thus eliminating the need for cutting the DNA. Consequently, these factors can either enhance or inhibit the expression of the gene.
A synergistic activation or repression effect is produced when natural gene expression regulation factors bind at multiple sites on a promoter. Cellecta’s dual-sgRNA CRISPRa and CRISPRi libraries are designed to enhance the activity of the standard single-sgRNA libraries since each construct expresses two different sgRNA binding distinct sites on the promoter of each gene target. As a result, it increases the likelihood of each target gene activating or repressing over a specified threshold in comparison to libraries where a single sgRNA targets a gene promoter.
Donato Tedesco, Ph.D., Director of R&D at Cellecta, said, “We found several examples where targeting more than one sgRNA to the same promoter enhanced expression levels of the target gene using the CRISPRa system, even when one sgRNA by itself had no detectable effect. As a result, it made sense to build a library where each construct has more than one sgRNA targeting each gene. We expected this to increase the overall level of effectiveness for the library, and indeed, this is what we saw when we compared the overall expression levels of our single-sgRNA CRISPRa library with the dual-sgRNA version.”
The new dual-sgRNA CRISPRa and CRISPRi lentiviral libraries make the gain-of-function and loss-of-function screens more efficient for researchers, while at the same time, producing better screening results. The offering by Cellecta provides a new range for scientists and researchers to aid in functional genetic screening, enabling accelerated identification of genome targets for therapeutics and biomarker analysis.