Functional and specific targeting for high-confidence gene knockout results
CRISPR-Cas9 is a highly effective tool for interrogating gene function, yet not all guide RNAs (gRNA) are effective in attaining functional protein knockout. To address this problem, Dharmacon developed an algorithm that is trained and validated for functional gene knockout, not just the ability to create insertions and deletions (indels). Edit-R pre-designed lentiviral single guide RNA (sgRNA) are CRISPR reagents for generating functional gene knockouts in difficult-to-transfect cells or for following up hits from a pooled lentiviral sgRNA library screen.
Edit-R Lentiviral sgRNAs express a chimeric structure comprised of a crRNA and tracrRNA fused through a short linker for the programming of Cas9 nuclease and creation of DNA double-strand breaks (DSBs). In the Edit-R lentiviral sgRNA vector backbone, the gene-specific crRNA and tracrRNA are expressed under the control of a human U6 promoter, while expression of the puromycin resistance marker (PuroR) is driven from the mouse CMV promoter and allows for rapid selection of cells with integrated sgRNA. Each Edit-R Lentiviral sgRNA is specific to the gene or genomic site of choice.
The Edit-R Lentiviral Gene Engineering system utilizes Cas9 nuclease and the single guide RNA in a two-step process. First, Edit-R Lentiviral Cas9 Nuclease Expression particles are utilized to generate cell lines stably expressing Cas9 nuclease. These cells can subsequently be transduced with Edit-R Lentiviral sgRNA particles to achieve efficient gene editing for phenotypic analyses in a population of cells or in isolated clonal cell lines.
100% of Edit-R predesigned guide RNA are guaranteed to edit!
Edit-R predesigned crRNA and lentiviral sgRNAs are guaranteed to edit your target, or we will replace it! No restrictions on Cas9 nuclease formats – if your Edit-R positive control works, so will your gene-specific guide RNA. Click on the Specifications tab to learn more about our guarantee.
Predesigned Edit-R Lentiviral sgRNAs are supplied as glycerol stocks and as concentrated high-titer particles for straightforward delivery into Cas9-expressing cells for efficient knockout without need for cloning or packaging. Purified, concentrated lentiviral particles can be directly transduced into difficult-to-transfect cells without the toxic cellular debris and contaminants found in supernatant preparations. Glycerol stocks can be grown and the expression plasmid purified for immediate transfection or packaging into particles.
Save money by eliminating time-consuming cloning, packaging and titering steps
High quality, concentrated, purified lentiviral particles for direct transduction with minimal cytotoxicity
Predesigned sgRNA generated using the Edit-R CRISPR RNA algorithm for unparalleled specificity and functionality
The Edit-R Predesigned Guide RNA Guarantee
We guarantee that EVERY predesigned Edit-R CRISPR-Cas9 crRNA and Edit-R lentiviral sgRNA (guide RNAs) will provide successful editing at the target site when delivered as described in the Edit-R Technical Manuals.
The Edit-R guide RNA guarantee is valid when used with any wild type S. pyogenes Cas9 nuclease, including mRNA, expression plasmid, protein, or stable Cas9 expression, and Edit-R crRNAs must be used with Edit-R tracrRNA for the guarantee to apply.
Analysis of editing of the treated cell population must be shown using a T7E1 or Surveyor mismatch detection assay. If successful editing is not observed for a predesigned Edit-R guide RNA while an appropriate side-by-side Edit-R positive control is successful, a one-time replacement of a different predesigned Edit-R guide RNA of the same format and quantity will be provided at no cost.
A replacement will only be approved upon discussion with our Technical Support team (firstname.lastname@example.org).
Successful editing at the DNA level does not always lead to functional gene knockout; it is recommended to test multiple guide RNAs to determine the most effective guide RNA for knockout of your target gene.
This guarantee does not extend to any accompanying experimental costs, does not apply to guide RNAs ordered via the CRISPR Design Tool, and will not be extended to the replacement guide RNA.
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