gRNA Expression Vectors
CRISPR (clustered regularly interspaced short palindromic repeats) and CRISPR-associated protein 9 (Cas9) are components of the leading genome or gene editing system, CRISPR-Cas9. Since it was developed in 2012, this gene editing tool has revolutionized biology research, making it easier to study disease and enabling faster drug discovery. The catalytically inactive Cas9 (dCas9) protein has greatly expanded this gene editing system to foster the study of gene expression and regulation in a wide range of organisms. This dCas9 protein lacks the ability to create DNA breaks in the target and serves as a DNA binding protein. When it fuses with a transcriptional repressor such as the Krüppel-associated box (KRAB) repressor or a transcriptional activator such as the tripartite fusion of three transcription activation domains: VP64, p65 and Rta (VPR), it can effectively regress or activate gene expression. To do this, a single guide RNA (sgRNA) of the gene of interest must be present. The Cas9-sgRNA complex binds to DNA sequences that are complementary to the sgRNA and causes a steric block that halts transcript elongation by RNA polymerase in CRISPRi systems. While in CRISPRa systems, the Cas9-sgRNA complex recruits transcription factors to increase desired gene expression.
Our CRISPRi and CRISPRa systems consist of two components in separate lentiviral vectors, the dCas9-KRAB for CRISPRi or the dCas9-VPR for CRISPRa vectors and the sgRNA expression vectors. Three sgRNA expression vectors are available at Alstem, with different selection markers BSD, PURO, and NEO (neomycin).
Gene specific gRNA that usually targets the promoter region is cloned into one of the sgRNA expression vectors in all the sgRNA vectors, human U6 promoter drives the sgRNA expression.
These two types of lentivirus vectors with different selection markers can be used together by co-transduction into the cell or to generate a stable dCas9-KRAB cell line by transduction with a gene-specific gRNA vector to achieve transcriptional inhibition of the target gene.
|ID||Catalog#||Name||Unit||Unit Price (USD)||Actions|
|289||CG101||pLV-U6-gRNA scaffold-EF1-BSD||10 μg at 0.5 μg/μL in TE||$695.00||Add to Cart|
|290||CG102||pLV-U6-gRNA scaffold-EF1-PURO||10 μg at 0.5 μg/μL in TE||$695.00||Add to Cart|
|291||CG103||pLV-U6-gRNA scaffold-EF1-NEO||10 μg at 0.5 μg/μL in TE||$695.00||Add to Cart|
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