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    Synthetic sgRNA

    Custom single guide RNA (sgRNA) for CRISPR-Cas9 gene editing

    These guide RNAs provide a high level of gene editing functionality and are provided as transfection-ready RNA oligos. This synthetic offering gives researchers a choice in guide RNAs, when a single molecule is preferred for therapeutic applications. Synthetic sgRNAs can be designed and ordered for S. pyogenes, or any other species of Cas9 with the CRISPR Design Tool.

    • Single chimeric molecule to simplify therapeutic development
    • Improved nuclease resistance with chemical modifications (2xMS)
    • Design guide RNAs for over 30 model organisms
    • Reduced immune response compared to in vitro transcribed sgRNAs

    Overview

    Edit-R CRISPR-Cas9 synthetic single guide RNAs greatly simplify the workflow of permanently knocking out genes by eliminating the time-consuming cloning of individual guide RNA expression vectors. Our chimeric sgRNAs contain both the crRNA and tracrRNA in one molecule that arrives ready-to-use and is ideal for experiments when a single molecule is preferable. Edit-R synthetic sgRNA provide the functionality and reliability of Dharmacon RNA oligos.

    Single guide RNAs can be designed to target any gene using the CRISPR Design Tool. The sgRNA is synthesized using patented Dharmacon 2’-ACE chemistry with faster and more efficient coupling rates than standard RNA synthesis chemistries resulting in more full length product. Synthetic sgRNA are column-purified, deprotected, and desalted to give highly functional, ready-to-use molecules.

    For larger quantities and additional purification options for sgRNAs the Custom single-strand RNA synthesis tool can be used for ordering.

    Synthetic sgRNA are compatible with any type of Cas9 nuclease, it is simply a matter of appropriate transfection conditions:

    Cas9 nuclease expression Recommended transfection reagent
    Stably expressed Cas9 nuclease DharmaFECT 1, 2, 3, or 4 for transfection of synthetic guide RNA
    Cas9 nuclease expression plasmid DharmaFECT Duo for co-transfection of plasmid and synthetic guide RNA
    Cas9 mRNA or protein NLS DharmaFECT Duo for co-transfection of mRNA or protein and synthetic guide RNA

    How many experiments can I do with the synthetic sgRNA?

    This table provides the approximate number of experiments that can be carried out for lipid transfection methods at the recommended sgRNA working concentration (25 nM) in various plate/well formats. Calculations do not account for pipetting errors.
    sgRNA
    nmol
    96-well plate
    100 µL reaction volume
    24-well plate
    500 µL reaction volume
    12-well plate
    1000 µL reaction volume
    6-well plate
    2500 µL reaction volume
    1 400 80 40 16

    Custom Guide RNA Design & Ordering Tools

    • 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.

    • Custom RNA synthesis

      Place a custom sgRNA order for larger scales and additional purification options.

    HDR Donor Template Design & Ordering Tools

    HDR Donor Template Kits

    Workflow

    Supporting Data

    Resources

    References

    1. M.L. Kelley, Ž. Strezoska, et al.Versatility of chemically synthesized guide RNAs for CRISPR-Cas9 genome editing. J. Biotechnol. 233, 74–83 (2016). doi:10.1016/j.jbiotec.2016.06.011
    2. Briner, A.E., Donohoue, P.D., et al.,Guide RNA functional modules direct Cas9 activity and orthogonality.Mol Cell. 56(2), 333-339 (2014).
    3. Hsu, P.D., Scott, D.A., et al.,DNA targeting specificity of RNA-guided Cas9 nucleases.Nat. Biotechnol. 31(9), 827-832 (2013).
    4. Hendel, A, et. al. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nat.Biotechnol. 33, 985-989 (2015).

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