Edit-R synthetic crRNA negative controls

Gene editing
Gene editing reagents
Edit-R Synthetic crRNA negative controls

Edit-R synthetic crRNA negative controls

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

Synthetic guide RNAs for efficient gene knockout & unparalleled specificity

Guaranteed to edit the target gene of interest
Designed to maximize the likelihood of functional protein knockout and minimize off-target editing
Transfection-ready RNAs eliminate cloning and in vitro transcription steps
Chemically modified for improved nuclease resistance

Reminder: Synthetic crRNA must be paired with synthetic tracrRNA to form the full guide RNA complex that targets Cas9 editing.

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 the Stabilizing modifications on Edit-R crRNA and tracrRNA article.

AAVS1 cutting controls are designed to target regions of the genome that do not result in functional knockout of a protein or known phenotypic changes. These regions are sometimes known as safe harbor regions. These guides are validated for mismatch detection assays and are recommended for screening applications as an additional control that results in Cas9 cutting without a phenotypic readout.

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.

Highlights

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!

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

Yes, these controls have the 2xMS modification pattern, as described in the Stabilizing modifications on Edit-R crRNA and tracrRNA blog article.

Our neutral cutting controls target an inactive region of the genome to provide a quantitative measure of cutting efficiency by mismatch detection assays, while maintaining a negative or negligible phenotype.

The neutral cutting controls target the AAVS1 safe harbor site on chromosome 19. This locus is a well-established and well-characterized inactive genomic region.

Our non-targeting control guide RNAs do not target any region in the genome. Non-targeting designs control for phenotypic changes that may be caused by the delivery of the CRISPR-Cas9 reagents in the absence of cutting. However, the non-targeting controls cannot indicate changes caused by Cas9 cutting because they do not result in cutting by the Cas9 enzyme.

Our positive controls target established housekeeping genes to offer insight into phenotypic changes that might be due to Cas9 cutting but are not specific to cutting within the gene of interest. However, there may be cell types or culture conditions under which cutting at the housekeeping gene locus could result in a phenotype, or experimental situations where disrupting a gene is not desired. To overcome this potentially confounding effect, the neutral cutting control can be included to provide more robust results and increased confidence in the phenotypes observed following editing of the target gene.

Neutral cutting controls can be helpful if there is concern that an observed phenotype is not gene-specific. In particular, neutral cutting controls are helpful as an additional control in screening applications and when a phenotype is observed with a positive control guide RNA. Additionally, this guide can be used in discovery settings where disruption of a gene could affect downstream phenotypes and results.