慢病毒表达载体pLV-SFFV-dCas9-VPR-PGK-

Gene Activation Vectors：CRISPR (clustered regularly interspaced short palindromic repeats) activation (CRISPRa) is a widely used genetic technique that allows for targeted activation of transcription in bacteria and mammalian cells. The CRISPRa system is comprised of a catalytically inactive Cas9 (dCas9) protein fused with a transcriptional activator such as the tripartite fusion of three transcription activation domains: VP64, p65 and Rta (VPR) and a customizable single guide RNA (sgRNA). The Cas9-sgRNA complex binds to DNA elements complementary to the sgRNA and recruits transcription factors to increase gene expression, resulting in the activation of the target gene.

Our dCas9-VPR system has two sets of separate lentiviral vectors: the dCas9-VPR vectors and the gRNA expression vectors. There are five individual versions of the dCas9-VPR expression vectors with either an EF1 or an SFFV promoter that drives the dCas9-VPR fusion protein expression and a PGK promoter that controls the expression of a selection marker, either puromycin (PURO) or Blasticidin S (BSD). The gRNA expression vectors consist of three versions with different selection markers BSD, PURO, and NEO (neomycin). Gene specific gRNA that usually targets the promoter region is cloned into one of the gRNA expression vectors. In all the sgRNA vectors, human U6 promoter is used to drive the sgRNA expression.

These two types of lentiviral vectors with different selection markers can be used together by co-transduction into the cells to achieve transcriptional activation of the target gene. Alternatively, a stable dCas9-VPR cell line can be generated and then transduced with a gene-specific gRNA vector to achieve the same purpose.
Description:CRISPR (clustered regularly interspaced short palindromic repeats) activation (CRISPRa) is a widely used genetic technique that allows for targeted activation of transcription in bacteria and mammalian cells. The CRISPRa system is comprised of a catalytically inactive Cas9 (dCas9) protein fused with a transcriptional activator such as the tripartite fusion of three transcription activation domains: VP64, p65 and Rta (VPR) and a customizable single guide RNA (sgRNA). The Cas9-sgRNA complex binds to DNA elements complementary to the sgRNA and recruits transcription factors to increase gene expression, resulting in the activation of the target gene.

pLV-SFFV-dCas9-VPR-PGK-BSD Lentiviral Vector expresses the dCas9-VPR fusion with a SFFV promoter and BSD with a PGK promoter to allow for selection of transduced cells.

Vector Information：This is a lentiviral expression vector that contains all the elements necessary for efficient and high yield viral production. In this vector, ubiquitous SFFV and PGK promoters drive the expression of the dCas9-VPR fusion protein and BSD, respectively, to allow for the selection of transduced cells. This vector can be used for stable cell line generation or in combination with a gene specifc sgRNA to activate the target gene expression.

IMPORTANT NOTICE

Store the vial at -20°C immediately upon receipt.

Specifications:

Product Name	pLV-SFFV-dCas9-VPR-PGK-BSD
Shipping Condition	Room Temperature - 2 Day Shipping
Storage and Stability	Store at -20°C immediately upon receipt. This product is stable for 6 months when stored as directed.
Quality Control	This plasmid is sequence verified.
Restricted Use	For Research Use Only. Not for use in diagnostic or therapeutic procedures.