CRISPRi synthetic sgRNA positive controls will selectively repress a well characterized human gene (either PPIB1 or SEL1L). Positive controls are recommended for optimization and ongoing monitoring of delivery conditions and CRISPRi dCas9-SALL1-SDS3 expression prior to experiments with target gene-specific guide RNAs.
- Select PPIB1 or SEL1L as your positive control gene
- Choose individual or pooled CRISPRi sgRNA controls to match your experimental CRISPRi sgRNA reagent format
- Ideal for optimization of experimental conditions and confirming successful gene repression
In addition to positive controls, it is recommended to use a CRISPRi synthetic sgRNA non-targeting control to determine baseline levels of gene expression.
Robust gene knockdown with CRISPRi synthetic sgRNA positive controls
Robust transcriptional gene repression with co-delivery of dCas9-SALL1-SDS3 mRNA and CRISPRi synthetic sgRNA positive controls. K562 and Jurkat cells were nucleofected with dCas9-KRAB or dCas9-SALL1-SDS3 mRNA (2 µg), and pooled CRISPRi synthetic sgRNAs (5 µM) targeting PPIB and SEL1L via a Lonza 96-well Shuttle system. WTC-11 hiPS cells were nucleofected with dCas9-KRAB or dCas9-SALL1-SDS3 mRNA (1 µg) and pooled CRISPRi synthetic sgRNA (3 µM) targeting PPPIB or SEL1L via a Lonza 96-well Shuttle system. Cells were harvested 72 hours post-nucleofection. Total RNA was isolated, and relative gene expression was measured using RT-qPCR. The relative gene expression for each target gene was calculated with the Cq method using GAPDH as the housekeeping gene and normalized to a non-targeting control (NTC).