CRISPRainbow Multiplex Labeling Kit
(Kit #
1000000079
)
Depositing Lab: Thoru Pederson
The CRISPRainbow technology developed by the Pederson lab uses CRISPR/Cas9 to tag up to seven different genomic locations. This unique labeling system will allow scientists to track DNA movement in live cells and study the structure of the genome in real time.
This kit will be sent as bacterial stabs.
Original Publication
Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T. Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. PubMed PMID: 27088723 .
Description
CRISPRainbow is a system that uses dead Cas9 (dCas9) and gRNA scaffolds that bind fluorescent proteins. This technique was used to simultaneously image six colors in order to label chromsomal loci.
To generate the various colors, different hairpins (MS2, PP7, or boxB) were inserted into stem loops of the gRNA. Colors were generated by recruiting pairs of fluorescent proteins (BFP, GFP, or RFP) that were fused to RNA hairpin binding domains:
| Color | Hairpin |
|---|---|
| Blue | MS2-MS2 |
| Green | PP7-PP7 |
| Red | boxB-boxB |
| Cyan | MS2-PP7 |
| Yellow | PP7-boxB |
| Magenta | boxB-MS2 |
| White | boxB-MS2-PP7 |
Labeling multiple chromsome-specific loci from Ma et al, 2016. PubMed PMID: 27088723 .
In the above figure, one CRISPRainbow color was used for one chromsomal locus:
- Blue (Cx): X chromosome
- Green (C14): chromosome 14
- Red (C7): chromosome 7
- Cyan (C1): chromosome 1
- Yellow (C13): chromosome 13
- Magenta (C3): chromosome 3
sgRNA design for tertiary color generation of white; Represents sgRNA with three elements (boxB, MS2, and PP7) as well as the three binding partners with their respective fluorescent proteins; from Ma et al, 2016. PubMed PMID: 27088723 .
Depositor-provided Files
UMass Medical School communication (includes article and video interview)
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The CRISPRainbow kit was a gift from Thoru Pederson (Addgene kit # 1000000079).”
For your Reference section:
Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T. Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. PubMed PMID: 27088723 .
Kit Contents
This kit will be sent as bacterial stabs.
Please note that plasmid pHAGE-EFS-PCP-3XNeon-nls is not included in this kit. If you need this plasmid for experiments, it must be ordered separately.
| ID | Name |
|---|---|
| 75381 | pHAGE-TO-dCas9 |
| 75383 | pHAGE-TO-MCP-3XBFPnls |
| 75384 | pHAGE-EFS-MCP-3XBFPnls |
| 75385 | pHAGE-EFS-PCP-3XGFPnls |
| 75387 | pHAGE-EFS-N22p-3XRFPnls |
| 75388 | pLH-sgRNA1 |
| 75389 | pLH-sgRNA1-2XMS2 |
| 75390 | pLH-sgRNA1-2XPP7 |
| 75391 | pLH-sgRNA1-2XboxB |
| 75392 | pLH-sgRNA1-MS2-PP7 |
| 75393 | pLH-sgRNA1-PP7-boxB |
| 75396 | pLH-sgRNA1-boxB-MS2 |
| 75397 | pLH-sgRNA1-boxB-MS2-PP7 |
| 75398 | pCRISPRainbow-DONOR1 |