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Single guide RNA powered by lentiviral vectors
Choose from our predesigned human, mouse, or rat sgRNA 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.
The Edit-R Lentiviral Gene Engineering platform includes two critical components based on the natural S. pyogenes system: a vector for Cas9 expression and a gene-specific vector for sgRNA expression designed to the target site of interest. Edit-R Lentiviral single guide RNA (sgRNA) reagents simplify the workflow of permanently knocking out genes by eliminating the time-consuming cloning and lentiviral packaging of individual guide RNA expression vectors. Our approach includes predesigned or custom ready-to-use sgRNA lentiviral particles which enable gene editing experiments in difficult-to-transfect cells.
In contrast to the two-RNA system of Edit-R synthetic crRNA and tracrRNA, the sgRNA expresses both as a single chimeric transcript.
Figure 1. Illustration of Cas9 nuclease programmed by the sgRNA complex cutting both strands of genomic DNA 5' of the PAM
Algorithm-optimized sgRNA for genome-wide coverage of human, mouse, or rat genes. Provided as high-titer lentiviral particles and glycerol stocks.
Species-specific sgRNAs targeting well-characterized genes 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 sgRNA
High-titer pooled screening libraries for pre-defined gene sets in human and mouse.
Arrayed collections of lentiviral sgRNA libraries for high-throughput knockout screening across entire human gene families.
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
Gene knockout workflow using the Edit-R Lentiviral Cas9 Nuclease with sgRNA system. Gene editing with Edit-R Lentiviral Cas9 Nuclease and sgRNA can be done following a mixed cell populations approach (left side) typically for gene knockout screenings or on isolated clonal cells lines (right side) when a defined genotype is desired or required on each step for the phenotypic analyses.
A recombinant U2OS ubiquitin-EGFP proteasome cell line (Ubi[G76V]-EGFP) and HEK293T cells were stably transduced with lentiviral particles containing Cas9 and a blasticidin resistance gene. A population of stably integrated cells were selected with blasticidin for a minimum of 10 days before transduction with sgRNAs. Cells were transduced with sgRNA lentiviral particles at low MOI to obtain cells with one integrant and selected with puromycin for seven days prior to analysis. The relative frequency of gene editing in the puromycin-selected cells was calculated from a DNA mismatch detection assay using T7 Endonuclease I.