Catalog #: CRUTEST

CRISPR-Test Cas9 and dCas9 Activity Test Kits

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Cellecta's CRISPR Functional Cas9 and dCas9 Activity Test Kits measure the functional activity of Cas9 nuclease, dCas9-repressor, or dCas9-activator in any vertebrate cell line.

1. CRISPRuTest™ Functional Cas9 Activity Assay Kit (Catalog # CRUTEST) can be used to measure the Cas9 nuclease activity in any vertebrate cell system. The CRISPR system uses a target single-guide RNA molecule (sgRNA) to target and cut a specific sequence on the genomic DNA with the Cas9 nuclease. The CRISPRuTest Kit provides a FACS-based assay to assess the Cas9 nuclease activity in cells expressing Streptococcus pyogenes Cas9. The kit contains pseudoviral-packaged lentiviral constructs with a green fluorescent protein (GFP) gene whose fluorescence is disrupted in cells with active Cas9. The kit provides enough reagents to measure this activity in 5-15 cell lines.

2. CRISPRaTest™ Functional dCas9-Activator Assay Kit (Catalog # CRATEST) measures functional dCas9 transactivator (e.g., dCas-VPH) in any mammalian cell system. The CRISPRaTest Kit provides reagents to measure the activator strength of any Streptococcus pyogenes dCas9 transactivator (dCas9-TA) hybrid protein in cells designed to be used for CRISPRa experiments using a FACS-based assay. The kit contains pseudoviral packaged lentiviral constructs with a green fluorescent protein (GFP) gene that increases in expression when expressed in cells with active dCas9-TA. The kit provides enough reagents to measure this activity in 5-15 cell lines.

3. CRISPRiTest™ Functional dCas9-Repressor Assay Kit (Catalog # CRITEST) allows you to measure a functional dCas9-Repressor (e.g., dCas9-KRAB) in any mammalian cell system. The CRISPRiTest Kit provides reagents to measure the repressor efficiency of any Streptococcus pyogenes dCas9 repressor (dCas9-Repressor) hybrid protein in cells designed to be used for CRISPRi experiments, using a FACS-based assay. The Kit contains packaged lentiviral constructs expressing dCas9-Rep regulated GFP and a non-regulated RFP protein. The Kit provides enough reagents for 5-10 cell lines.

Combination kits --including CRISPR-Test Functional Cas9 or dCas9 Assay bundled with Cas9/dCas9 with 4 different promoters-- are available at a reduced price when purchased in combination, as compared to purchasing the items separately. These are useful for determining the optimal Cas9 or dCas9 repressor or activator promoter for your cell system.

a. CRISPRuTest™ Cas9 Assay with 10^6 TU of each of the following 4 constructs (Catalog #: CRUTP-SET)

    • pRCCB-CMV-Cas9-2A-Blast
    • pRCE2B-EFS-Cas9-2A-Blast
    • pRCRB-RSV-Cas9-2A-Blast
    • pRCSFB-SFFV-Cas9-2A-Blast

b. CRISPRaTest™ dCas9-Activator Assay with 10^6 TU of each of the following 4 constructs (Catalog #: CRATP-SET)

    • pRDVCCB-CMV-dCas9-VPH-2A-Blast
    • pRDVCE2B-EFS-dCas9-VPH-2A-Blast
    • pRDVCRB-RSV-dCas9-VPH-2A-Blast
    • pRDVCSFB-SFFV-dCas9-VPH-2A-Blast

c. CRISPRiTest™ Cas9 Assay with 10^6 TU of each of the following 4 constructs (Catalog #: CRITP-SET)

    • pRDKCCB-CMV-dCas9-KRAB-2A-Blast
    • pRDKCE2B-EFS-dCas9-KRAB-2A-Blast
    • pRDKCRB-RSV-dCas9-KRAB-2A-Blast
    • pRDKCSFB-SFFV-dCas9-KRAB-2A-Blast

An alternative to measuring functional Cas9 or dCas9 activity by measuring loss of GFP is to use the CRISPRtest Essential-Gene Cas9 Assay, which relies on cell death induced by disruption of a gene required for viability.

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