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Description：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 detection of 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.
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. pLV-U6-gRNA scaffold-EF1-NEO sgRNA Lentiviral Vector expresses gRNA with the human U6 promoter and NEO with the EF1 promoter to allow for NEO selection of transduced cells.
Vector Information：This is a lentiviral expression vector that contains all the elements for efficient and high yield viral production. A ubiquitous EF1 promoter drives the expression of NEO to allow for the selection of transduced cells. This vector is used in combination with the Cas9-KRAB or the Cas9-VPR to repress or activate the target gene expression, respectively.
IMPORTANT NOTICE：Store the vial at -20°C immediately upon receipt.
Note: Bacterial culture of pLV vectors should be done in medium containing 10 μg/mL Ampicillin. For maximal plasmid yield and quality, we recommend Stbl3 competent cells (Invitrogen). Specifications:
| Product Name | pLV-U6-gRNA scaffold-EF1-NEO |
|---|---|
| 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. |
