CRISPRi for transcriptional repression
CRISPR interference (CRISPRi) allows researchers to down-regulate specific gene function by blocking transcription, without editing the DNA. Visit our CRISPRi applications page to learn more.
The system requires two components to operate: a gene-specific guide RNA and a catalytically deactivated Cas9 (dCas9) fused to transcriptional repressor domains (SALL1 and SDS3).
Predesigned CRISPRi guide RNA
Algorithm-designed lentiviral and synthetic single guide RNA (sgRNA) specifically target immediately downstream of the gene's transcriptional start site (TSS).
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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.
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CRISPRi lentiviral sgRNA
An ideal delivery method for CRISPRi in difficult-to-transfect cells, or when longer term repression is desired.
CRISPRi dCas9-SALL1-SDS3
Nuclease-deactivated Cas9, fused to our proprietary transcriptional repressor domains SALL1 and SDS3.
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CRISPRi dCas9-SALL1-SDS3 mRNA
Co-transfect with synthetic sgRNA using Dharmafect transfection reagents for transient CRISPRi in lentiviral-free workflows.
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CRISPRi dCas9-SALL1-SDS3 lentiviral particles
Create a stable dCas9 expressing cell line, ready for CRISPRi studies.
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 dCas9-SALL1-SDS3 source | Guide RNA format | Delivery method | Benefits & Recommended uses |
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CRISPRi dCas9-SALL1-SDS3 lentiviral particles transduction & selection for CRISPRi dCas9 stable cells |
CRISPRi synthetic sgRNA | Transfection or electroporation |
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CRISPRi lentiviral sgRNA particles | Transduction |
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CRISPRi dCas9-SALL1-SDS3 mRNA | CRISPRi synthetic sgRNA | Co-transfection or electroporation |
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CRISPRi synthetic guide RNA
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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.
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CRISPRi synthetic sgRNA positive controls
Control sgRNAs targeting well-characterized genes to determine the effectiveness of your experimental conditions
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CRISPRi synthetic sgRNA non-targeting controls
Non-targeting controls to evaluate baseline cellular responses to CRISPRi components in the absence of target-specific crRNA
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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
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CRISPRi lentiviral sgRNA
Lentiviral particles of vector-based single guide RNA are predesigned for genome-wide coverage of human genes
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CRISPRi lentiviral sgRNA positive controls
Control sgRNAs targeting well-characterized genes to determine the effectiveness of your experimental conditions
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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).
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CRISPRi dCas9-SALL1-SDS3 mRNA
Co-transfect with synthetic sgRNA using Dharmafect transfection reagents for transient dCas9-SALL1-SDS3 expression in lentiviral-free workflows.
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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.