CRISPRa (Transcriptional Activation) system is a powerful tool for activating the transcription of endogenous genes. Mutated Cas9 (dCas9) connects with the transcriptional activator (such as VP64, p65 and HSF1) can effectively promote the transcription of specific genes. The gene transcription can be increased by two to thousands folds.

▍Vitro Biotech’s CRISPRa Service Features

1. Three vectors system, which makes transduction more efficient;
2. Screen for the sgRNA that has the most desired activation effect;
3. It acts directly in the promoter and is not limited to some certain transcripts;
4. It is an ideal choice for big-size gene overexpression;
5. Stable and long-term overexpression.

CRISPRa stable cell line workflow

Vitro Biotech will design gRNAs according to the target gene, construct the vectors, and package the lentivirus to infect target cells. The highly expressed or appropriate gRNA is screened, and then generate the CRISPRa stable cell line, and use RT-qPCR to validate the expression level of the target gene. We also provide single cell cloning and phenotypic assays upon request.

▍  CRISPRa vs ORF overexpression


CRISPRa
 ORF overexpression
Expression level
 Different gRNAs have different activation level
 Stable at certain expression level
Expression specificity
 Highly specific, affected by endogenous promoter
 Highly specific
ORF length limit
 No
 Yes, the longer the ORF, the lower the transfection efficiency
Expression characteristic
 As same as the isoforms of the target gene in the target cells
 Depending on the ORF