Description
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
