Gene Editing Reagent Core (GERC)

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Director: Dr. Lisette Maddison



Summary:

Tyrosinase CRISPR mutants test Directed by Dr. Lisette Maddison, the Gene Editing Reagent Core (GERC) provides a fee based service to provide CRISPR/Cas9 reagents for investigators to incorporate cutting edge genome editing into their research. The mission of the core is to help investigators integrate cutting edge gene editing into their research. The GERC provides expertise in design and implementation of CRISPR/Cas9 and reagents will be developed in close consultation with the investigators for personalized results. Available services include guide RNA and Cas9 supplied as independent reagents or as plasmid vectors, and development of repair templates for precise genomic changes from small to large.

Services and Costs:


Service

Cost Range

RNP - sgRNA and Cas9 Protein$179.00 - $195.00
RNP - sgRNA and Cas9 Protein (w/validation in cell line)$468.00 - $508.00
Expression Constructs - U6 driven sgRNA, CBH Promoter Cas9
-dual vs sgRNA and separate Cas9 vector
$248.00 - $269.00
Expression Constructs - U6 driven sgRNA, CBH Promoter Cas9
-dual vs sgRNA and separate Cas9 vector (w/validation in cell line)
$549.00 - $597.00
Targeting oligo for single/small bp changes$244.00 - $266.00
Targeting vectors for large changes$2,702.00 - $3592.00
(subject to complexity)

Please contact Dr. Maddison with any projects requests not addressed above.

More Information:

To tailor services for the individual goals of each PI, the GERC director will first meet with the investigator to discuss the goals of the project(s) and the desired reagents.

The core is currently positioned to provide the following services:

  1. Development of gene specific guide RNA for genome mutation.
    • CRISPR/Cas9 is a customizable two part system where the gene specific element (guide RNA or gRNA) targets the desired gene or genomic location and the Cas9 protein cuts the DNA. Repair of this lesion can lead to deletions or insertions that disrupt the targeted genes. These repairs are random and require downstream analysis to identify those with the most desired outcome which can then be studied further.
    • The GERC will provide customized reagents for this outcome in two available formats depending on investigator's interest.
      1. The gRNA provided as in vitro synthesized RNA where the crRNA and the tracrRNA is produced as a single molecule. Cas9 can be provided as either protein or as RNA.
        • The Cas9 protein and gRNA together forms the ribonucleoprotein (RNP) which is sufficient to induce lesions in cells or embryos.
        • This may be a preferred reagent for investigators who solely desire to develop a stable mutation for downstream studies.
      2. The gRNA and Cas9 cloned into a plasmid vector. These can either be as dual vectors where gRNA and Cas9 are contained in a single plasmid or gRNA and Cas9 can be provided in separate plasmids.
  2. Development of repair template.
    • If the investigator desires a precise change in the genomic DNA or incorporation of elements such as fluorescent molecules, a template containing the desired changes has to be supplied alongside the Cas9 and gRNA. Depending on the complexity of the changes desired these can be provided in two different forms, both of which will be available through the GERC. These templates have to be carefully designed in order to achieve the desired outcome.
      1. A single stranded DNA template for small changes such as a single base pair change or incorporation of small epitope tags.
      2. A double stranded template for larger changes or incorporation of large elements. The GERC is positioned to efficiently construct these templates with a collection of available elements and expertise in rapid assembly of complex vectors.
Three images of embryos at the top, left two are Tyrosinase CRISPR mutants with pigmentation, right image is the wild-type. Below is an image showing the mutation analysis with the wild-type allele show on the right side.

CONTACT:

Dr. Lisette Maddison
509-335-8992
lmaddison@vetmed.wsu.edu

 



Washington State University