The most widely used application of CRISPR-Cas9 is to target a gene of interest and generate a double-strand break (DSB) at the target site. But what if you could use the highly specific CRISPR-Cas9 system to target your gene of interest and alter its expression without cutting the DNA backbone? Making use of a deactivated or dead Cas protein bound to activator or repressor domains achieves just that. These systems are used to alter the expression of gene(s) of interest in either gain- (CRISPR activation, or CRISPRa) or loss-of-function (CRISPR interference, or CRISPRi) experiments. Adding new dimensions to CRISPR, these systems are used to affect expression at the transcriptional level for novel insights into gene function.
While similarities do exist between CRISPRi and CRISPR knockout for loss-of-function experiments, CRISPRi allows for multiplexed repression experiments without concern for DSBs triggering cell death via the DNA damage response pathway. In contrast to traditional gene overexpression techniques, such as complementary DNA (cDNA) and open reading frames (ORFs), both of which produce excessive amounts of protein in the cell, CRISPRa provides a more biologically relevant overexpression model by upregulating genes within their native context. Used in parallel with other methods, CRISPRa and CRISPRi ensure more robust datasets through orthogonal validation.
During the webinar, the viewers will:
- Discover how CRISPR modulation (CRISPRmod) differs from CRISPR-Cas9 gene editing
- Learn the basics of CRISPRa and CRISPRi methodology
- Obtain insights into the benefits and key applications of CRISPRa and CRISPRi