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Site-specific Chromatin Loop Formation by dCas9

CLOuD9 Gene Expression Regulation System: Study the dynamic effects of chromatin structure on gene expression
• Site-specific chromatin loops
• Inducible, reversible loop formation
• Easy CRISPR/Cas9 targeting
• Well-validated system

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Form chromatin loops with CLOuD9 in an inducible and reversible way!

The CLOuD9 System consists of three components:

1. A null mutant of the Cas9 protein from S. aureus (sa-dCas9) fused to the plant ABI
protein (10 ug plasmid DNA are provided in the kit)
2. A null mutant of the Cas9 protein from S. pylogenes (sp-dCas9) fused to the plant
PYL1 protein (10 ug plasmid DNA are provided in the kit)
3. Inducer reagent: The plant phytohormone S-(+)-abscisic acid (ABA) (50 ul 1000x ABA
are provided in the kit. The reagent is also available separately in a 200 ul pack size.)

The use of the Cas9 null mutants enables accurate binding to a specific region of the
genome via the user-designed guide RNA (gRNA) without generating any breaks in the
DNA. Because the gRNA scaffolds from the two species bind more efficiently to their
same-species Cas9 partner, saCas9-ABI fusion localizes to one region of the DNA and the
spCas9-PYL1 fusion to a second region of DNA.

The dimerization partners—ABI and PYL1—are part of the plant ABA signaling pathway,
and form heterodimers in the presence of ABA. Dimerization of ABI and PYL1 is
reversible, enabling formation of chromatin loops in the presence of ABA, and
disassembly of the chromatin loops upon ABA washout.


Explore a new frontier in the study of gene expression regulation

Available only from SBI and featured in a recent Nature Communications article (see link
below), the CLOuD9 Gene Expression Regulation Kit gives you an exciting and innovative
tool for studying—and potentially controlling—gene expression regulation by putting you
in charge of chromatin loop formation. Gain a clearer understanding of the dynamic
effects of chromatin restructuring on the expression of your gene-of-interest and
increase gene expression in certain cellular contexts. Combining the simplicity of
CRISPR/Cas9 targeting—free from DNA cleavage activity—with inducible and reversible
protein dimerization partners, the CLOuD9 Gene Expression Regulation Kit adds a new
dimension to your gene regulation studies.

CLOuD9 for Chromatin Loop Reorganization using CRISPR-dCas9:

• Innovative—leverage the simplicity of CRISPR/Cas9 targeting—free of DNA cleavage
activity—for inducible and reversible chromatin loop formation anywhere in the genome
• Well-validated—chromatin loop formation and the resulting changes in gene expression
across multiple loci and cell lines is amply demonstrated in Morgan, et al (see link below)
• Broadly applicable—great for studying the role of chromatin in the control of gene
expression in a range of areas, including:
- Developmental biology
- Cancer biology
- Chromatin structural biology
- Chromatin-based therapeutic strategies
- Unique—the innovative CLOuD9 System is only manufactured by SBI

The CLOuD9 Gene Expression Regulation Kit comes with CLOuD9 Inducer Reagent and
the two lentivector plasmids needed for expressing CLOuD9’s CRISPR/Cas9 System
components:

One lentivector plasmid expressing a null Staphylococcus aureus Cas9 protein fused to
one of the protein dimerization partners—the ABI protein from the plant abscisic acid
(ABA) signaling pathway

A second lentivector plasmid expressing a null S. pyogenes Cas9 protein fused to the
other protein dimerization partner—the PYL1 protein from the plant ABA signaling
pathway

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Related Links

User Manual
Reference Paper in Nature Communications (Morgan et al.)
Validation Data

 

 

Description Cat# Size Price    
CLOuD9 Gene Expression Regulation Kit: Site-specific chromatin loop formation by dCas9 CASCL9-100A-KIT-SBI 1 kit 1101 € DETAILS   Add to Cart 
Inducer Reagent (1000x) for CLOuD9 Gene Expression Regulation (Plant phytohormone ABA) CASCL9-200A-1-SBI 200 ul 384 € DETAILS   Add to Cart 

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