Description
The binding of Programmed Cell Death Protein 1 (PD-1), a receptor expressed on activated T-cells, to its ligands, PD-L1 and PD-L2, negatively regulates immune responses. PD-1 ligands are found on most cancers, and the PD-1:PD-L1/2 interaction inhibits T-cell activity and enables cancer cells to escape immune surveillance. The PD-1:PD-L1/2 pathway is also involved in regulating autoimmune responses, making these proteins promising therapeutic targets for a number of cancers, as well as multiple sclerosis, arthritis, lupus, and type I diabetes.
This sgRNA (single guide RNA) pool is designed for use with the CRISPR Synergistic Activation Mediator (SAM) system to induce transcriptional activation and expression of any gene of interest. CRISPRa (SAM) cells stably express a mutated dCas9 (Streptococcus pyogenes CRISPR associated protein 9), lacking any endonuclease activity, fused with VP64, a transcriptional activator, as well as transcription factors P65 (Transcription Factor p65, or Nuclear Factor NF-κB p65) and HSF1 (Heat Shock Factor 1) fused with an MS2 tag.
When these cells are transfected with an MS2-tagged sgRNA targeting the promoter region of PD-1 (Programmed Cell Death 1, PDCD1, CD279, GenBank Accession #NM_005018), dCas9-VP64 and MS2-P65-HSF1 are recruited to the site in the genomic DNA and begin transcription, inducing PD-1 expression. Each pool includes 5 individually validated sgRNA to ensure robust expression