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CRISPR-Cas9 Gene Editing

Optimized tools for high-confidence genome engineering

The Dharmacon Edit-R CRISPR-Cas9 platform greatly simplifies the workflow of permanently knocking out genes. Our approach includes predesigned, ready-to-use DNA and RNA components and enables fast assessment of multiple target sites per gene for multiple genes.

Explore our products for successful CRISPR-Cas9 genome engineering

  • CRISPR Guide RNA

    High quality, ready-to-use lentiviral and synthetic reagents to guide Cas9 cleavage

  • Cas9 Nuclease

    Configure the optimal promoter for your cell type to ensure robust Cas9 expression or explore DNA-free options

  • CRISPR Controls and Detection Primers

    Proper controls are essential to assessment of CRISPR-Cas9 genomic editing experiments

  • CRISPR-Cas9 Screening Libraries

    Pooled sgRNA or arrayed crRNA for high-throughput gene editing studies

  • HDR Donor Templates

    Synthetic single-stranded DNA oligonucleotide (ssDNA oligo) donors for precise genomic modification with the homology-directed repair (HDR) pathway

Introduction to CRISPR

The ability to precisely and permanently alter endogenous gene expression through targeted genome editing is a highly effective reverse genetics tool. Recently, genome engineering has advanced tremendously with the characterization of bacterial and archael CRISPR (clustered regularly interspaced short palindromic repeats) systems and their adapted usage in mammalian cells. (Figure 1)

CRISPR systems have been described in the literature as innate immune defense systems analagous to eukaryotic RNA interference (RNAi) pathways. Additionally, the Cas9 (CRISPR-associated 9) nuclease has been defined as a dedicated effector enzyme that cleaves DNA when guided by two required small RNA sequences: the CRISPR RNA (crRNA) which binds the target DNA and guides cleavage, and the trans-activating RNA (tracrRNA) which base-pairs with the crRNA and enables the Cas9-crRNA complex to locate the targeted DNA. Recent publications demonstrate this system can be engineered to target and cleave DNA in mammalian cells, thereby permanently disrupting gene expression, making this system a new and exciting molecular tool to interrogate gene function.

Recommended Reading

Below is a selection of important journal articles in the CRISPR-Cas9 research field.

  1. R. Barrangou, A. Birmingham et. al. Advances in CRISPR-Cas9 genome engineering: lessons learned from RNA interference. Nucleic Acids Res. 43(7), 3407-3419 (2015)
  2. D. Bhaya, M. Davison, et al. CRISPR-Cas systems in bacteria and archaea: versatile small RNAs for adaptive defense and regulation. Annu. Rev. Genet. 45, 273-297 (2011).
  3. L. Cong, F. A. Ran, et al. Multiplex Genome Engineering Using CRISPR/Cas Systems. Science. 339(6121), 819-823 (2013).
  4. E. Deltcheva, K. Chylinski, et al. CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. Nature. 471(7340), 602-607 (2011).
  5. Y. Fu, J. D. Sander, et al. Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat. Biotechnol. (2014).
  6. D.Y. Guschin, A. J. Waite, et al. A rapid and general assay for monitoring endogenous gene modification. Methods Mol. Biol. 649, 247-256 (2010).
  7. F. Heigwer, G. Kerr, et al. E-CRISP: fast CRISPR target site identification. Nat. Methods. 11(2), 122-123 (2014).
  8. P.D. Hsu, D. A. Scott, et al. DNA targeting specificity of RNA-guided Cas9 nucleases. Nat. Biotechnol. 31(9), 827-832 (2013).
  9. M. Jinek, K. Chylinski, et al. A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 337(6096), 816-821 (2012).
  10. P. Mali, L. Yang, et al. RNA-guided human genome engineering via Cas9. Science. 339(6121), 823-826 (2013).
  11. N. K. Pyzocha, F. A. Ran, et al. RNA-Guided Genome Editing of Mammalian Cells. Methods Mol. Biol. 1114, 269-277 (2014).
  12. D. Reyon, C. Khayter, et al. Engineering designer transcription activator-like effector nucleases (TALENs) by REAL or REAL-Fast assembly. Curr. Protoc. Mol. Biol. 100, 12.15.1‐12.15.14 (2012).
  13. T. R. Sampson, D. S. Weiss. Exploiting CRISPR/Cas systems for biotechnology. Bioessays. 36(1), 34-38 (2014).
  14. T. Wang, J. J. Wei, et al. Genetic screens in human cells using the CRISPR-Cas9 system. Science. 343(6166), 80-84 (2014).

CRISPR-Cas9 Gene Editing Products

Synthetic Guide RNA

  • Edit-R predesigned crRNA

    Algorithm-optimized crRNA for genome-wide coverage of human, mouse, or rat genes. Simply search for your gene!

  • Edit-R tracrRNA

    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.

  • Edit-R Synthetic Positive crRNA Controls and Detection Primers

    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.

  • Edit-R Synthetic crRNA Non-targeting Controls

    Non-targeting controls to evaluate cellular responses to CRISPR-Cas9 components in the absence of gene target-specific crRNA.

  • Edit-R crRNA Libraries

    Plated pre-defined collections of popular human and mouse gene families for arrayed knockout screening.

  • Cherry-pick libraries

    Customize and order plates of predesigned crRNA for knockout studies for your targets of interest.

Lentiviral Guide RNA

Vector-based Cas9 Nuclease

DNA-free Cas9 Nuclease

Custom Guide RNA Ordering & Design Tool

  • CRISPR RNA Configurator

    Place a custom order, or design and order your own crRNA or lentiviral sgRNA with our easy-to-use interface.

  • Edit-R HDR Donor Designer

    Use this tool to design and order a single-stranded DNA donor (≤ 150 nt) for precise CRISPR-Cas9 gene editing with the HDR pathway.

The breadth of the Dharmacon portfolio supports multiple gene editing workflows; choose the one that suits your experimental system and desired outcome.

CRISPR-Cas9 Gene Editing Resources

CRISPR RNA Configurator

Do you have a favorite guide RNA sequence?

Do you require design assistance to ensure Cas9 cleavage in a specific region of your target gene?

Edit-R predesigned crRNA and lentiviral sgRNA provide algorithm-optimized designs for functionality and specificity; but when particular designs or parameters are desired, you can design your own crRNA or sgRNA with our easy-to-use interface.

The Dharmacon CRISPR RNA Configurator allows you to design or enter your own CRISPR targeting sequences for incorporation into synthetic crRNAs and single guide RNAs (sgRNAs).

See the CRISPR RNA Configurator User Guide for detailed instructions.