MilliporeSigmas Introduces CRISPRi Whole-Genome Libraries and Pools

CRISPRi modulates gene expression rather than imprecise DNA repair, allowing for robust loss-of-function studies, including in essential genes, with less off-target effects.

FREMONT, CA: MilliporeSigma, a leading provider of functional genomics screening and genome-editing software, has introduced whole-genome CRISPR Inhibition (CRISPRi) libraries and pools to support its industry-leading suite of genome-editing products and technologies. CRISPRi may be used to scan an entire genome, a small gene family, or a single target. The CRISPRi libraries are completely customizable, offering the same versatility as MilliporeSigma's entire family of screening items.

CRISPRi modulates gene expression rather than imprecise DNA repair, allowing for robust loss-of-function studies, including in essential genes, with less off-target effects. When only gene knockout or overexpression experiments are used, CRISPRi offers valuable insights into the underlying biology that are often overlooked. Researchers may make scientific breakthroughs by using CRISPRi as a development method to find new disease mechanisms or drug targets.

The entire CRISPRi libraries and pools were developed by the University of California, San Francisco, and then improved by MilliporeSigma, resulting in superior algorithm design and improved scaffolding that has been shown to increase gene knockdown, even in difficult-to- repress targets. The CRISPRi packages include:

• Pooled libraries holding the top five guide RNAs (gRNA) and a subpool of five additional gRNAs per gene, for a total of ten gRNAs targeting each gene; this provides improved sensitivity to most competitive technologies, which only deliver three to five gRNAs.

• Druggable genome, membrane proteins, cancer and apoptosis, stress and proteostasis genes, and others are among the gene groups targeted by established subpanels.

• A proprietary expression-stabilizing UCOE feature is included in a KRAB-dCas9 helper build.

• For the easier downstream study, CRISPRi vectors with blue fluorescent protein and puromycin selection markers were engineered.

• 10x Genomics-compatibility for downstream single-cell analysis.

CRISPR activation (CRISPRa) whole-genome libraries and pools, which are commonly used in combination with CRISPRi, are also part of MilliporeSigmas extensive gene modulation offering.

CRISPR knockout, RNAi knockdown, and open reading frame gain-of-function studies are among the company's screening methods for genome-editing applications. Genome editing and gene regulation are complementary technologies that, when combined, can fully elucidate gene pathways and recognize drug targets. MilliporeSigma is a global leader in developing groundbreaking CRISPR methods, products, and services to support every phase of genome engineering-facilitated research, from basic research to therapeutic delivery, with 17 years of experience in the area.