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 or mouse 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 and mouse 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.
- Proprietary alignment tools used to verify at least three mismatches or gaps to any potential target in human or mouse 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:
- A plasmid, protein, mRNA, or lentiviral vector expressing a mammalian codon-optimized gene sequence encoding Cas9 nuclease
- A chemically synthesized trans-activating CRISPR RNA (tracrRNA), and
- A chemically synthesized CRISPR RNA (crRNA) designed to the gene target site of interest
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|
Workflow for co-transfection of Cas9 nuclease plasmid and synthetic crRNA:tracrRNA
- R. Barrangou, A. Birmingham, et. al. Advances in CRISPR-Cas9 genome engineering: lessons learned from RNA interference. Nucleic Acids Research, 43(7) 3407-3419 (2015)
- 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