Edit-R predesigned synthetic sgRNA

Gene editing
Gene editing reagents
Edit-R predesigned synthetic sgRNA

Edit-R predesigned synthetic sgRNA

Synthetic single guide RNAs for efficient gene knockout & unparalleled specificity

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

Edit-R predesigned synthetic sgRNA

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Functional and specific targeting for high-confidence gene knockout results

Edit-R predesigned synthetic sgRNAs are chemically synthesized guide RNA for S. pyogenes Cas9 nuclease targeting and creation of Cas9-mediated DNA double-strand breaks in a genomic region of interest.

All Edit-R guides are algorithmically designed to efficiently generate functional protein knockouts, not just create an insertion or deletion. Additionally, chemical modifications are applied to all Edit-R synthetic guide RNA products to reduce degradation by nucleases and improve overall editing performance.

This unmatched accuracy and efficiency makes Edit-R predesigned synthetic sgRNA an ideal tool for pathway analysis or following up hits from a synthetic sgRNA library screen.

New! Human functionality and specificity scores now available.

All guide RNA designs are top algorithm picks for each gene; qualitative ranks for functionality and specificity allow you to fine tune human guide RNA choice to your specific application. The functionality score is a predicted indication of how likely this guide is to produce a functional knockout. The specificity score is based on the predicted risk of cutting activity at potential off-target sites. To learn more, visit our algorithm for Edit-R guide RNA page

Components for CRISPR gene editing experiment using synthetic sgRNA

Edit-R synthetic sgRNA designed to target the gene of interest — choose from individual guides or sets of 3 guides for greater likelihood of functional knockdown
A source of Cas9 nuclease — choose from protein, mRNA, or lentivirus formats, or a stable Cas9 expressing cell line
Appropriate positive and non-targeting controls
DharmaFECT transfection reagent or electroporation for guide delivery

Need to scale up your experiments? Choose from arrayed collections of predesigned synthetic sgRNA libraries for screening across the human genome and gene families, or design your own custom mini-library with our Cherry-Pick library tool.

Complete your knockout experiment by adding these supporting reagents to your order

Cas9 nuclease options

Cas9 mRNA or Cas9 protein NLS can be co-transfected with Edit-R synthetic sgRNA for a fully DNA-free workflow.
Stable Cas9 expressing cell lines are ready to be transfected with Edit-R synthetic sgRNA for rapid gene knockout.

Synthetic guide RNA controls

Non-targeting controls are used to evaluate cellular responses to CRISPR-Cas9 components in the absence of gene target-specific sgRNA. Choose from 5 scrambled sequences guaranteed not to target any gene in the human or mouse genomes.
Positive controls targeting well-characterized genes are used to determine the effectiveness of your gene editing conditions for maximal efficiency. Choose from PPIB or DNMT3B.

Transfection reagents

DharmaFECT transfection reagents are optimized for improved delivery and reduced toxicity. The optimal DharmaFECT reagent for your experiment will depend on your Cas9 nuclease source and your cell type.

Cas9 nuclease source	Recommended transfection reagent
Stable Cas9 expressing cell line or Cas9 nuclease lentiviral particles	DharmaFECT 1,2,3, or 4 for transfection of synthetic guide RNA. Use this guide to find the recommended DharmaFECT reagent for your cell type.
Cas9 mRNA or protein NLS	DharmaFECT Duo for co-transfection of mRNA or protein and synthetic guide RNA

How much sgRNA do I need?

This table provides the approximate number of experiments that can be carried out for lipid transfection methods at the recommended sgRNA working concentration (25nM) in various plate/well formats. Calculations do not account for pipetting errors.

sgRNA 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	800	160	80	32
5	2000	400	200	80
10	4000	800	400	160

Flow cytometry analysis of protein knockout in CD4⁺ T cells using a single predesigned synthetic sgRNA per gene target

Primary human CD4⁺ T cells were nucleofected with Cas9 RNP using an individual predesigned synthetic sgRNA per gene target or a non-targeting control (NTC), via a Lonza 96-well Shuttle system. After 72 hours, functional knockout of CD28, CD3E, and CXCR3 were assessed as a percent of cells not expressing the target gene by FACS analysis. Cells were stained and normalized for CD4 expression using an Alexa Fluor 488 conjugated antibody and compared to each target CD28, CD3E, and CXCR3 using APC conjugated primary antibodies.

Functional knockout of FUS gene with a pool of synthetic sgRNA

Functional knockout of FUS was assessed in U2OS cells constitutively expressing Cas9 under the CAG promoter. Cells were transfected with a 25 nM sgRNA pool containing three different Edit-R predesigned sgRNAs targeting FUS using 0.04 µL DharmaFECT 4. 72 hours post transfection the cells were split, and at 96 hours post transfection, cells were fixed and stained with a primary antibody targeting FUS, and an Alexa Fluor 488 conjugated fluorescent secondary antibody. Hoechst stain was used to identify nuclei.