Human CRISPRi Control and Effector Kit，1KIT，CRISPRICON-1KT，Sigma

General description

The control kit includes validated positive control targeting RAB1A and non-targeting control for assay set-up and a UCOE KRAB-dCas9 for consistent effector expression across a wide variety of cell lines. The kit is ideal for setting up the screen assay and optimization before using the CRISPRi library pools.

Application

• Set up and optimization of screen assay
• Functional Genomics/Target Validation
• Creation of cell lines stably expressing KRAB-dCas9
• Validated Positive and negative controls

Biochem/physiol Actions

The power of CRISPR for genome engineering, coupled with the ability to perform large-scale, whole genome, loss-of-function (LOF) screening, has allowed for new breakthroughs identifying gene pathways in drug resistance and disease. CRISPR is most commonly used to create double-stranded breaks that often result in loss of gene function (CRISPR-KO). However, the full extent of CRISPR′s utility extends beyond just targeted cutting of DNA. Nuclease-independent applications of CRISPR provide equal targeting specificity but instead of cutting, CRISPRi allows for targeted interference of gene function by delivering transcriptional repressor domains to a specific target sequence using modified dCas9 + gRNA complexes. Gene knockdown is complementary to CRISPR-KO and CRISPRa (activation) and has distinct advantages over existing loss-of-function strategies like RNAi. We partnered with University of California San Francisco to provide the best-in-class CRISPRi screening tools available.

Features and Benefits

• Ease of optimization: Validated positive control targeting RAB1A and non-targeting control for assay set up
• Best in Class UCSF gRNA selection algorithm and optimized (F+E) gRNA scaffold
• UCOE KRAB-dCas9 for consistent effector expression across a wide variety of cell lines
• Libraries provided at high functional titer, based on FACs or CFU analysis
• Use puromycin and or BFP as selection markers

Other Notes

2 Controls and 1 Effector Construct with a minimum concentration of 5x10⁵ TU/mL(via FACS or CFU assay)

CRISPRIE: KRAB-dCas9 - Blasticidin CRISPRi Construct Lentiviral Transduction Particles
CRISPRI01: CRISPRi RAB1A Puromycin Control Transduction Particles
CRISPRI06: CRISPRi Nontargeting Control Puromycin Transduction Particles

For more information about screening with CRISPRi/a please Click Here.

Legal Information

CRISPR Use License Agreement

Lentiviral, WPRE and Evrogen Fluorophore Licenses