CRISPR guide RNA

Confidence in your knockout experiment

Dharmacon Edit-R CRISPR guide RNAs are guaranteed to edit the target gene-of-interest, so you can be confident in your knockout result – see guarantee tab for details.

Our guide RNAs are designed using a validated algorithm to achieve functional gene knockout with high specificity. By assessing phenotypes for thousands of designs, then validating our design rules in multiple assay systems, we have established standards for determining optimal knockout target sites.

Visit our application page to learn more about the Edit-R CRISPR-Cas9 gene editing system.

Flexibility to fit your workflow

We offer Dharmacon Edit-R CRISPR guide RNA in pools or as individual reagents in a variety of formats to fit many different experimental workflows.

Synthetic single guide (sgRNA) or two part crRNA:tracrRNA reagents can be co-transfected with Cas9 mRNA or protein for DNA-free workflows.
Lentiviral sgRNA are recommended for editing in difficult-to-transfect cells.
All-in-one lentiviral sgRNA + Cas9 offers a single-reagent knockout workflow.
Custom design synthetic or lentiviral guide RNAs for additional species, alternative nucleases or any target location using the CRISPR Design Tool.

Scale to meet any scope

Expand the scope of your next loss-of-function experiment. Use the Cherry-Pick Library Tool to design and order your own custom CRISPR knockout screening library of Edit-R guide RNA reagents. Or see our predefined CRISPR knockout libraries for gene families or whole genome screens.

Order CRISPR guide RNA

Synthetic sgRNA

Transfection-ready synthetic sgRNAs eliminate cloning and in vitro transcription steps

Synthetic crRNA

Must be used with synthetic tracrRNA

All-in-one sgRNA + Cas9

Stable guide RNA expression for cell types not amenable to transfection

Lentiviral sgRNA

Stable guide RNA expression for cell types not amenable to transfection

Custom guide RNA solutions

CRISPR Design Tool

Design and order custom guide RNA for any species, nuclease or cut site

Custom guide RNA

When you already know your guide RNA sequence and need to get it synthesized

Guide RNA reagent selection

Determining the most appropriate guide RNA format for your experiment will depend on your particular application and cell type. Use this table to select the right reagents for your specific experimental conditions.

	Synthetic crRNA	Synthetic sgRNA	Lentiviral sgRNA	Lentiviral All-in-one sgRNA
Guaranteed to edit your gene of interest	✔	✔	✔	✔
Can be custom designed for any species, nuclease or editing location	✔	✔	✔*	✔*
RNP compatible for electroporation	✔	✔
Transduce into difficult-to-edit cells that can't be electroporated			✔	✔
Population enrichment with antibiotic resistance markers			✔	✔
Population enrichment with fluorescent markers	✔**	✔**		✔
Single reagent workflow				✔

*custom vector designs for use with S. pyogenes Cas9 only.
**when co-transfected with fluorescent Cas9 mRNA.

Edit-R algorithm scores correlate to cleavage efficiency — 10 crRNAs with high functional scores for 10 genes (blue bars) and 10 crRNAs with low functional scores for the same genes (yellow bars) were tested for editing by Next Generation Sequencing. 93% of the high-scoring crRNAs and 32 % of the low scoring crRNAs showed > 40% of editing (indel formation). The Cas9-HEK293T cell line was transfected with 50 nM crRNA:tracrRNA, using 0.25 µL/well of DharmaFECT 1. Seventy-two hours post-transfection, cells were lysed and Nextera transposon-adapted amplicons spanning each crRNA site were generated for every treated sample as well as for a matched control amplicon from untransfected samples. Samples were indexed using the Nextera 96-well index kit and pooled for sequencing on a MiSeq instrument (paired end reads, 2 x 300 length). Reads that passed NGS quality filtering criteria were aligned to the reference file (Bowtie2 v2.1.0). Percent perfect reads were calculated and normalized to the control untransfected samples (Samtools v0.1.12a); the data is presented as normalized percent edited.

crRNAs with high algorithm functional scores also perform well in other functional assays — U2OS-Proteasome cells with integrated Cas9 (under CAG promoter) were plated in 96-well plates at 10,000 cells per well. 24h after plating, cells were transfected with 25 nM crRNA:tracrRNA using 0.2 µg/well of DharmaFECT4. Cells were analyzed for apoptosis 48 h after transfection using the ApoONE homogeneous caspase assay (Promega). Shown is a box plot representation of the functionality of crRNAs targeting BCL2L1, PLK1 or WEE1 in ApoONE assay. To generate the box plot, the crRNAs were divided into bottom half (H1) and top half (h2) based on their functional score. The medians, distribution of data between the lower and upper quartile and the minimum and maximum values demonstrate that high-scoring crRNAs have increased functionality.

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.

CRISPR guide RNA

Dharmacon Edit-R synthetic guide RNA & controls

Synthetic sgRNA

Genome wide synthetic sgRNA designs guaranteed to edit your gene of interest. The design algorithm maximizes the potential for functional protein knockout while minimizing off-target editing through stringent specificity checks.
- Synthetic sgRNA positive controls and detection primers
  
  Species-specific synthetic sgRNAs targeting well-characterized genes, as well as mismatch detection assay primers, to determine the effectiveness of your gene editing conditions for maximal efficiency.
- Synthetic sgRNA non-targeting controls
  
  Non-targeting controls to evaluate cellular responses to CRISPR-Cas9 components in the absence of gene target-specific sgRNA.
- Cutting controls
  
  Cutting control (safe harbor) synthetic sgRNAs recommended for determination of baseline cellular responses in CRISPR-Cas9 experiments.

Synthetic crRNA

Guaranteed to edit your target! Algorithm-optimized crRNA for genome-wide coverage of human and mouse genes. Modifications for nuclease resistance improve DNA-free editing.
Synthetic tracrRNA

Trans-activating CRISPR RNA (tracrRNA) is required for use with Edit-R synthetic crRNA to form the complex that programs Cas9 nuclease.

Dharmacon Edit-R lentiviral sgRNA & controls

All-in-one lentiviral sgRNA + Cas9

Combined Cas9 and sgRNA expression for efficient gene knockout & unparalleled specificity; available as glycerol stocks and high-titer purified particles.

All-in-one lentiviral sgRNA positive controls and kits

All-in-one lentiviral sgRNA controls to verify DNA double-strand breaks and gene editing efficiencies.
All-in-one lentiviral sgRNA non-targeting controls

All-in-one lentiviral sgRNA constructs bioinformatically designed to not target any gene in human or mouse genomes.

Lentiviral sgRNA

Guaranteed to edit your target, algorithm-optimized sgRNA for genome-wide coverage of human or mouse genes. Provided as high-titer lentiviral particles or glycerol stocks.

Lentiviral sgRNA positive controls

Species-specific sgRNAs targeting well-characterized genes to determine the effectiveness of your gene editing conditions for maximum efficiency.
Lentiviral sgRNA non-targeting controls

Non-targeting controls to evaluate cellular responses to CRISPR-Cas9 components in the absence of gene-specific sgRNA.

Custom guide RNA design

CRISPR Design Tool

Place a custom guide RNA order, or design and order your own synthetic sgRNA, crRNA, or lentiviral sgRNA with our easy-to-use interface.

Dharmacon Edit-R HDR donor template design, ordering tools & kits

HDR Donor Designer - oligo

Design and order a single-strand DNA donor (≤ 150 nt) for insertion, deletion, or other alteration.
HDR Donor Designer - plasmid

Design and order a plasmid DNA donor kit for insertion of a mKate2 or EGFP fluorescent marker, or other a custom insert.
HDR plasmid donor kits

Rapidly and easily assemble a plasmid donor for HDR
HDR plasmid donor components

Quickly and efficiently build a HDR donor plasmid
HDR plasmid donor primers

PCR components for Edit-R Plasmid Donor Kits

Cas9 nuclease sources

"CRISPR-ready" premade Cas9-expressing cell lines

Cas9 expressing stable cell lines

Choose from a variety of popular cell backgrounds, then simply deliver a Edit-R CRISPR guide RNA targeted to any gene for simple loss-of-function studies.

Transient Cas9 nuclease for DNA-free workflows

Cas9 nuclease mRNA

Purified Cas9 mRNA for transient Cas9 nuclease expression. Fluorescent options available for sorting, enrichment and visualization of delivery.
Cas9 nuclease protein NLS

Purified Cas9 protein ready-to-use for DNA-free workflows.

Vector-based solutions for generating Cas9-expressing cell lines

Lentiviral Cas9 nuclease reagents

Purified lentiviral particles or plasmid DNA for generation of stable Cas9 nuclease-expressing cell populations. Constitutive or inducible promoter options are available.

CRISPR guide RNA

Algorithm designed CRISPR guide RNAs for highly accurate and efficient functional gene knockout

Confidence in your knockout experiment

Flexibility to fit your workflow

Scale to meet any scope

Order CRISPR guide RNA

Custom guide RNA solutions

Guide RNA reagent selection

Supporting data

Edit-R algorithm scores correlate to cleavage efficiency

crRNAs with high algorithm functional scores also perform well in other functional assays

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

Reagents & controls

CRISPR guide RNA

Dharmacon Edit-R synthetic guide RNA & controls

Cas9 nuclease sources

"CRISPR-ready" premade Cas9-expressing cell lines

Screening libraries

Guarantee

The Dharmacon Edit-R guarantee