CRISPRa lentiviral sgRNA non-targeting controls are recommended as negative controls for experiments using CRISPRa lentiviral sgRNAs in human and mouse cells. They are designed to have a minimum of three mismatches or gaps to all potential PAM-adjacent targets in human, mouse and rat genomes. When using these controls, changes in cellular viability or gene expression likely reflect nonspecific cellular responses that can be used as a baseline for comparison to cells treated with target-specific reagents.
- CRISPRa guide RNA non-targeting control sequences are cloned into the same optimized lentiviral vector as CRISPRa gene targeting sgRNAs
- Proprietary alignment tools used to verify at least three mismatches or gaps to any potential target in human, mouse or rat genomes
- Available as high-titer purified lentiviral particles or glycerol stocks
CRISPRa workflow diagram with stable dCas9-VPR expression
CRISPR activation workflow with lentiviral dCas9-VPR and synthetic crRNA:tracrRNA (left side) or Lentiviral expressed sgRNA (right side).
CRISPRa plasmid co-transfection workflow
CRISPRa workflow for plasmid co-transfection of lentiviral dCas9-VPR and expressed sgRNA.
Transductions with lentiviral sgRNA particles in different dCas9-VPR cell lines
U2OS, HEK293T, MCF 10A and K562 stably expressing integrated CRISPRa dCas9-VPR were plated at 10,000 cells/well and transduced with CRISPRa sgRNA lentiviral particles targeting POU5F1 or TTN at a MOI of 0.3 to obtain cells with a single integrant. Cells were selected with 2 µg/mL puromycin for 4 days prior to analysis with RT-qPCR. The relative expression of each gene was calculated with the Cq method using GAPDH as the housekeeping gene and normalized to a non-targeting control.
Certificate of analysis
Genome-wide human and mouse lentiviral sgRNA reagents are specifically designed for CRISPRa. Available as glycerol stocks and high-titer purified particles. When used in conjunction with dCas9-VPR expression, you can leverage the power of the CRISPR-Cas9 system for activation of your favorite gene.