Previously added items:
Pooled sgRNA or arrayed crRNA for high-throughput gene editing studies
Investigation of entire gene families or biological pathways is enabled by our broad suite of custom and pre-defined libraries of CRISPR-Cas9 reagents. Both pooled and arrayed screens can take advantage of our algorithm-optimized guide RNA designs for high functionality and superior specificity.
The Dharmacon Edit-R CRISPR-Cas9 platform includes predesigned, ready-to-use guide RNAs to enable rapid assessment of multiple targets across many genes. The application of this platform is greatly expanded by our offering of arrayed crRNA libraries and pooled lentiviral libraries. Both products enable unbiased, powerful loss-of-function screens.
Plated pre-defined collections of popular human and mouse gene families for arrayed knockout screening
Customize and order plates of predesigned crRNA for knockout studies for your targets of interest
The Edit-R trans-activating CRISPR RNA (tracrRNA) is a chemically synthesized and HPLC-purified long RNA required for use with synthetic crRNA to form the complex that programs Cas9 nuclease.
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.
When choosing a CRISPR-Cas9 screening platform, there are many considerations in determining the best tools for your experimental needs. Instrumentation, analytical support, and assay type(s) must all be taken into account.
Successful screening with CRISPR-Cas9 reagents is highly dependent on the efficiency of gene editing achieved. Both the Edit-R crRNA Libraries and Pooled sgRNA Screening Libraries benefit from the Edit-R algorithm, which selects for highly functional designs.
U2OS-(Ubi)EGFP-Cas9 stable cells were seeded at 5,000 cells/well in 96-well format. At 24 hours cells were transfected with 25 nM crRNA:tracrRNA complexes targeting the PLK1 and KIF11 and non-targeting controls (NTC) using 0.2 µl /well DharmaFECT4. Four different gene-targeting and NTC guide sequences were utilized and experimental controls included untransfected cells (UT) and cells treated with lipid alone. At 48 hours post-transfection, the phenotype was analyzed on the IN Cell Analyzer 2200 imaging system (GE Healthcare).
Upper panel: Mitotic Index (MI) was analyzed as percent cells positive for Phospho-Histone H3 (PH3) and was normalized to the average MI of the four negative NTC crRNAs.
Lower panel: Representative fluorescent micrograph showing the nuclei in blue and PH3 in red.
Here we are targeting the gene PSMD11 at 12 different sites and measuring functionality by EGFP signal to indicate disruption of the proteasome. Cells were either transfected with crRNA:tracrRNAs, or transduced with lentiviral particles for expression of a sgRNA of the same design. There is no significant difference between the two guide RNA formats when targeting the same genomic site; a good design for crRNA will translate into a good design for sgRNA.