CRISPRi assay considerations for reagent selection

There are many options and considerations for CRISPRi experimental conditions. Generally, users achieve the most robust knockdown when working with a stable population of dCas9-SALL1-SDS3 expressing cells. For extended timepoint assays (more than 120 hours), many choose lentiviral sgRNA. For short-term assays (less than 120 hours), synthetic sgRNA gives more robust gene repression than expressed guide RNAs. 

CRISPRi synthetic guide RNA

• CRISPRi synthetic sgRNA

  The most rapid CRISPRi system, with repression within 24 hours post-transfection. Synthetic RNAs are modified for nuclease resistance and available as pooled or individual reagents.

• CRISPRi synthetic sgRNA positive controls

  Control sgRNAs targeting well-characterized genes to determine the effectiveness of your experimental conditions

• CRISPRi synthetic sgRNA non-targeting controls

  Non-targeting controls to evaluate baseline cellular responses to CRISPRi components in the absence of target-specific crRNA 

• CRISPRi synthetic sgRNA cherry-pick libraries

  Upload your own gene list to customize and order plates of predesigned sgRNA for CRISPRi studies across tens or thousands of genes

CRISPRi lentiviral guide RNA

• CRISPRi lentiviral sgRNA

  Lentiviral particles of vector-based single guide RNA are predesigned for genome-wide coverage of human genes

• CRISPRi lentiviral sgRNA positive controls

  Control sgRNAs targeting well-characterized genes to determine the effectiveness of your experimental conditions

• CRISPRi lentiviral sgRNA non-targeting controls

  Non-targeting controls to evaluate baseline cellular responses to CRISPRi components in the absence of target-specific sgRNA

CRISPRi dCas9-SALL1-SDS3

Nuclease-deactivated Cas9, fused to our proprietary transcriptional repressor domains (SALL1 and SDS3).

• CRISPRi dCas9-SALL1-SDS3 mRNA

  Co-transfect with synthetic sgRNA using Dharmafect transfection reagents for transient dCas9-SALL1-SDS3 expression in lentiviral-free workflows.

• CRISPRi dCas9-SALL1-SDS3 lentiviral particles

  Create a stable dCas9-SALL1-SDS3 expressing cell line, ready for CRISPRi studies.

CRISPRi products may be combined in several ways to support your particular experimental needs. Find workflows below for 1) co-transfection with synthetic sgRNA and dCas9-SALL1-SDS3 mRNA, 2) using CRISPRi dCas9-SALL1-SDS3 lentiviral particles to create a stable dCas9-SALL1-SDS3 expressing cell line and introducing synthetic sgRNA, or 3) creating a stable cell line and introducing either plasmid or lentiviral sgRNA

CRISPRi workflow using synthetic sgRNA and dCas9-SALL1-SDS3 mRNA

Co-transfect or electroporate cells with CRISPRi synthetic sgRNA and CRISPRi dCas9-SALL1-SDS3 mRNA. Then enrich cell populations using fluorescence or puromycin resistance options. This system is best suited for rapid, transient gene repression studies.

CRISPRi workflow using synthetic sgRNA and dCas9-SALL1-SDS3 lentiviral particles

Transduce cells with dCas9-SALL1-SDS3 lentiviral particles to generate CRISPRi-ready, stable dCas9-SALL1-SDS3 expressing cells. Then introduce CRISPRi synthetic sgRNA specific to your target gene(s). This system is ideal for arrayed screening in transfectable cell models.

CRISPRi workflow using lentiviral sgRNA and dCas9-SALL1-SDS3 lentiviral particles

Workflow using CRISPRi dCas9-SALL1-SDS3 lentiviral particles to establish a stable dCas9-SALL1-SDS3 expressing cell line, then delivering CRISPRi lentiviral sgRNA particles (left) or plasmid sgRNA (right). This system is ideal when working with difficult-to-transfect cell types.