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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.

Add to Cart
$375 USD + shipping

Available to academics and nonprofits only.

CRISPR Rainbow color diagram

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
Fluorescent cell labeling using CRISPR rainbow

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 cartoon for white color

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.

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
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