Edit-R Synthetic crRNA Non-targeting Controls

Bioinformatically designed to not target any gene in human, mouse or rat genomes


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Non-targeting control crRNAs recommended for determination of baseline cellular responses in CRISPR-Cas9 experiments.

**Please note** our synthetic crRNA control catalog numbers have changed to reflect the addition of stabilizing chemical modifications to the crRNA for nuclease resistance. Learn more about these changes in this featured article.

Edit-R Synthetic crRNA Non-targeting Controls are recommended as negative controls for experiments using crRNAs in human, mouse, or rat cells. All Edit-R Non-targeting Controls are designed to have a minimum of three mismatches or gaps to all potential PAM-adjacent targets in human, mouse and rat genomes. Changes in viability or gene expression levels in cells treated with these controls likely reflect a baseline cellular response that can be compared to the levels in cells treated with target-specific crRNAs.

Highlights

  • Proprietary alignment tools used to verify at least three mismatches or gaps to any potential target in human, mouse or rat genomes
  • Five different designs to choose from, ensuring your chance of finding one that has no detectable effects in your system

Remember to order tracrRNA for use with your controls!

Edit-R CRISPR-Cas9 Gene Engineering Platform

The Dharmacon Edit-R Gene Engineering platform is based on the Type II CRISPR-Cas9 system from the bacteria Streptococcus pyogenes which can be engineered and adapted to edit genes in mammalian cells. When Cas9, the endonuclease component of a CRISPR-Cas system, is complexed with two RNAs called the CRISPR RNA (crRNA) and the trans-activating crRNA (tracrRNA), it forms a complex that cleaves DNA. This flexible system can be exploited to induce site-specific genome modifications to program, regulate and precisely interrogate gene function.

The Dharmacon Edit-R CRISPR-Cas9 platform includes the three critical components required for gene editing in mammalian cells, based on the natural S. pyogenes system:

 

How much crRNA & tracrRNA do I need?

This table provides the approximate number of experiments that can be carried out for lipid transfection methods at the recommended crRNA:tracrRNA working concentration (25 nM:25nM) in various plate/well formats. Calculations do not account for pipetting errors.
crRNA
nmol
tracrRNA
nmol
96-well plate
100 µL reaction volume
24-well plate
500 µL reaction volume
12-well plate
1000 µL reaction volume
6-well plate
2500 µL reaction volume
2 2 800 160 80 32
5 5 2000 400 200 80
10 10 4000 800 400 160
20 20 8000 1600 800 320
  
CategoryName
HazardousNo
Hazardous:No
Shelf Life:
Shipping ConditionAmbient
Shipping Condition:
Shipping Information
SpecificationName
SpecificationValue
Stability at Recommended Storage ConditionsAt least 12 months
Storage Condition-20 C
Storage Condition:
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Workflow for co-transfection of Cas9 nuclease plasmid and synthetic crRNA:tracrRNA

co-transfection-crispr-cas9-workflow.png


References

  1. R. Barrangou, A. Birmingham, et. al. Advances in CRISPR-Cas9 genome engineering: lessons learned from RNA interference. Nucleic Acids Research43(7) 3407-3419 (2015)
  2. M.L. Kelley, M.L., Ž. Strezoska, et al. Versatility of chemically synthesized guide RNAs for CRISPR-Cas9 genome editing. J. Biotechnol. 233, 74–83 (2016). doi:10.1016/j.jbiotec.2016.06.011 

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