Lentiviral sgRNA constructs bioinformatically designed and validated to not target any gene in human or mouse genomes
Negative control CRISPRa sgRNAs to establish experimental baselines and to distinguish sequence-specific biological effects from non-specific effects. Available as high-titer purified lentiviral particle and glycerol stock formats
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.