Previously added items:
Pre-designed or custom designs to power rapid, efficient gene editing
Choose from our predesigned human, mouse, or rat crRNA products to take advantage of our proprietary Edit-R algorithm to improve functionality and specificity, or design your own guide RNA to meet your particular needs.
Edit-R CRISPR-Cas9 synthetic reagents greatly simplify the workflow of permanently knocking out genes by eliminating the time-consuming cloning of individual guide RNA expression vectors. Our novel approach includes predesigned or custom ready-to-use reagents which enable fast assessment of multiple target sites per gene, for multiple genes.
In contrast to a single guide RNA which is expresses both the crRNA and the tracrRNA as a single chimeric transcript, the Edit-R synthetic crRNA and tracrRNA are transfected together to complex with the Cas9 nuclease to carry out DSB.
Synthetic crRNA:tracrRNA are compatible with any type of Cas9 nuclease, it is simply a matter of appropriate transfection conditions:
Algorithm-optimized crRNA for genome-wide coverage of human, mouse, or rat genes. Modifications for nuclease resistance improve DNA-free editing. Simply search for your gene!
Edit-R trans-activating CRISPR RNA (tracrRNA) is synthetic, HPLC-purified, long RNA required for use with Edit-R crRNA to form the complex that programs Cas9 nuclease. It is modified for nuclease resistance and can be used with modified or unmodified Edit-R crRNA.
Species-specific crRNAs targeting well-characterized genes, as well as mismatch detection assay primers, to determine the effectiveness of your gene editing conditions for maximal efficiency.
Non-targeting controls to evaluate cellular responses to CRISPR-Cas9 components in the absence of gene target-specific crRNA.
Plated pre-defined collections of popular gene families for arrayed knockout screening
Have a favorite gene list? Customize and order plates of predesigned crRNA for knockout studies in your targets of interest
Place a custom guide RNA order, or design and order your own synthetic sgRNA, crRNA, or lentiviral sgRNA with our easy-to-use interface.
Design and order a single-strand DNA donor (≤ 150 nt) for insertion, deletion, or other alteration.
Design and order a plasmid DNA donor kit for insertion of an mKate2 or EGFP fluorescent marker, or a custom insert
Rapidly and easily assemble a plasmid donor for HDR
Quickly and efficiently build a HDR donor plasmid
PCR components for Edit-R Plasmid Donor Kits
Figure 1. Edit-R gene engineering workflow using the Edit-R Cas9 Nuclease Expression plasmids co-transfected with synthetic crRNA and tracrRNA.
Figure 2. Gene knockout workflow using the Edit-R Lentiviral Cas9 Nuclease Expression particles with synthetic crRNA and tracrRNA.
To facilitate rapid generation of cell lines that constitutively express Cas9 nuclease, the Edit-R Lentiviral Cas9 Nuclease Expression vector is packaged into particles, purified and concentrated for direct transduction. Subsequent transfection of synthetic crRNA and tracrRNA into Cas9-expressing cell lines results in a higher percentage of edited cells in comparison to co-transfection of Cas9 plasmid DNA with synthetic crRNA and tracrRNA without enrichment.
Targeted DNA cleavage is achieved using Edit-R Cas9 Nuclease Expression plasmid programmed with crRNA:tracrRNA in HEK293T cells. HEK293T cells were transfected with Edit-R Cas9 Nuclease Expression Plasmid alone (- lanes) or with Edit-R Cas9 plus tracrRNA and crRNA targeting DNMT3B, PPIB or CDKN1A (+ lanes) using DharmaFECT Duo Transfection reagent. No enrichment was carried out. Direct lysis of cells was performed 72 hours post-transfection and PCR was performed with primers flanking the cleavage sites. A DNA mismatch assay with T7Endonuclease I (NEB) was performed and the samples were separated on a 2% agarose gel. Percent indels was calculated using ImageJ software and is shown at the bottom of the lanes.
A recombinant U2OS ubiquitin-EGFP proteasome cell line (Ubi[G76V]-EGFP) was stably transduced with lentiviral particles containing Edit-R plasmid vectors expressing Cas9 nuclease and blasticidin resistance gene under the control of the indicated promoters. A population of cells with stably integrated Cas9-BlastR was selected with blasticidin-treatment for a minimum of 10 days before transfections. Cells were transfected with 50 nM Edit-R synthetic crRNA:tracrRNA complex targeting a proteasome component,VCP, using DharmaFECT 3 Transfection Reagent. After 72 hours, transfected cells were examined for EGFP+ cells (upper panel) and the relative frequency of gene editing was calculated (lower panel) based on a DNA mismatch detection assay with T7 Endonuclease I.