Use of the CRISPRiTest assay requires access to a flow cytometer with the following excitation and emission requirements:
Excitation: 488nm and 561nm
Emission: 530/20nm and 590/20nm
Cellecta also offers the following products that may be useful when running this assay:
dCas9-Only Plasmids
| Cat.# | Description | Quantity |
|---|---|---|
| SVKRABC9B-PS | CRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Blast (plasmid) | 25 µg |
| SVKRABC9B-VS | CRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Blast (virus) | 1 × 106 TU |
| SVKRABC9H-PS | CRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Hygro (plasmid) | 25 µg |
| SVKRABC9H-VS | CRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Hygro (virus) | 1 × 106 TU |
| SVKRABC9B-PS | CRISPRi dCas9-KRAB pRDKCCBla-CMV-dCas9-KRAB-2A-Blast (plasmid) | 25 µg |
| SVKRABC9B-VS | CRISPRi dCas9-KRAB pRDKCCBla-CMV-dCas9-KRAB-2A-Blast (virus) | 1 × 106 TU |
| SVKRABC9G-PS | CRISPRi dCas9-KRAB pRDKCCG-CMV-dCas9-KRAB-2A-TagGFP2 (plasmid) | 25 µg |
| SVKRABC9G-VS | CRISPRi dCas9-KRAB pRDKCCG-CMV-dCas9-KRAB-2A-TagGFP2 (virus) | 1 × 106 TU |
| SVKRABC9H-PS | CRISPRi dCas9-KRAB pRDKCCH-CMV-dCas9-KRAB-2A-Hygro (plasmid) | 25 µg |
| SVKRABC9H-VS | CRISPRi dCas9-KRAB pRDKCCH-CMV-dCas9-KRAB-2A-Hygro (virus) | 1 × 106 TU |
| SVKRABC9N-PS | CRISPRi dCas9-KRAB pRDKCCN-CMV-dCas9-KRAB-2A-Neo (plasmid) | 25 µg |
| SVKRABC9N-VS | CRISPRi dCas9-KRAB pRDKCCN-CMV-dCas9-KRAB-2A-Neo (virus) | 1 × 106 TU |
| SVKRABC9R-PS | CRISPRi dCas9-KRAB pRDKCCR-CMV-dCas9-KRAB-2A-TagRFP (plasmid) | 25 µg |
| SVKRABC9R-VS | CRISPRi dCas9-KRAB pRDKCCR-CMV-dCas9-KRAB-2A-TagRFP (virus) | 1 × 106 TU |
| SVKRABC9EB-PS | CRISPRi dCas9-KRAB pRDKCEBla-EF1-dCas9-KRAB-2A-Blast (plasmid) | 25 µg |
| SVKRABC9EB-VS | CRISPRi dCas9-KRAB pRDKCEBla-EF1-dCas9-KRAB-2A-Blast (virus) | 1 × 106 TU |
| SVKRABC9ER-PS | CRISPRi dCas9-KRAB pRDKCER-EF1-dCas9-KRAB-2A-TagRFP (plasmid) | 25 µg |
| SVKRABC9ER-VS | CRISPRi dCas9-KRAB pRDKCER-EF1-dCas9-KRAB-2A-TagRFP (virus) | 1 × 106 TU |
| SVKRABC9E2B-PS | CRISPRi dCas9-KRAB pRDKCE2Bla-EFS-dCas9-KRAB-2A-Blast (plasmid) | 25 µg |
| SVKRABC9E2B-VS | CRISPRi dCas9-KRAB pRDKCE2Bla-EFS-dCas9-KRAB-2A-Blast (virus) | 1 × 106 TU |
| SVKRABC9RB-PS | CRISPRi dCas9-KRAB pRDKCRBla-RSV-dCas9-KRAB-2A-Blast (plasmid) | 25 µg |
| SVKRABC9RB-VS | CRISPRi dCas9-KRAB pRDKCRBla-RSV-dCas9-KRAB-2A-Blast (virus) | 1 × 106 TU |
Cell Line
Parental dCas9-KRAB+ MDA-MB-231 Cells (Cat.# ZMDAMB231-CKRA, 106 cells) may be run in parallel with the user’s dCas9-repressor (dCas9-Rep) cells as a positive control for the assay.
Transduction Control Virus
Non-Targeting CRISPR Control pRSG16-U6-sg-UbiC-TagRFP-2A-Puro (virus) may be used to assess transduction efficiency of the target cells.
| Cat.# | Description | Quantity |
|---|---|---|
| SGCTL-NT-pRSG16-V | Non-Targeting CRISPR control pRSG16-U6-sg-UbiC-TagRFP-2A-Puro Vector (virus) | 1 × 107 TU |
Non-targeting controls in other Cellecta vectors are available. Please visit our CRISPR Non-Targeting sgRNA Control Construct in Standard Lentiviral Vector web page.
Other than the specific reagents and instruments above, the protocol assumes the user has access to standard materials (e.g., polypropylene tubes, pipette tips), equipment (table top centrifuges, pipettes, scales), and common reagents (e.g., TE buffer, ethanol) and buffers used in a typical life science laboratory.
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