sgRNA clone for CRISPRi
Repress gene expression, without cutting the DNA
| Vector | Promoter | CRISPR option | Selection Marker / Reporter Gene | Vector Type | Vector Map |
|---|---|---|---|---|---|
| pCRISPR-AB04 | U6 | CRISPRi dCas9-KRAB | Puro / mCherry | Lentiviral |  |
Overview
The CRISPRi system comprises 2 components
dCas9-KARB and sgRNA were co-overexpressed in HeLa cells. The transcriptional activity of DPH2 was assessed by qPCR. The amplification curves for cells transduced with sgRNA are shown in red, while those for cells without sgRNA transduction are shown in blue. A comparison of the transcriptional levels of the target gene with those of GAPDH demonstrated that the transcriptional levels of the target gene were significantly suppressed in HeLa cells.
The CRISPRi system comprises 2 components
- dCas9-KRAB fusion protein. Guided by the sgRNA, the dCas9 protein binds to the target DNA. Its fused KRAB domain then recruits epigenetic enzymes, such as the histone deacetylases HDAC1/2 and the histone methyltransferases, by binding to co-repressor proteins, such as KAP1. This process represses the transcriptional level of the target gene.
- sgRNA
Application
dCas9-KARB and sgRNA were co-overexpressed in HeLa cells. The transcriptional activity of DPH2 was assessed by qPCR. The amplification curves for cells transduced with sgRNA are shown in red, while those for cells without sgRNA transduction are shown in blue. A comparison of the transcriptional levels of the target gene with those of GAPDH demonstrated that the transcriptional levels of the target gene were significantly suppressed in HeLa cells.