How to Make a Mouse Model using CRISPR with Emory’s Transgenic Mouse Core Facility
PPMS
Sigma
form
Sigma
Quote
Week 1
Oligo/
Plasmid
Crisprs
Delivered
Injection
Week 9
Week 10
Genotype
Toe
Week 14
Confirm +
Genotype
Transfer
Founders
Week 15
Week 18
Quick Guide (see below for more detailed flow chart):
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PI submits a project request form in PPMS
PI submits Crispr order form to Sigma and consults with Sigma on Crispr design and construction
Sigma produces and validates Crispr constructs and provides oligo or donor DNA sequence for plasmid (if applicable)
If project requires plasmid: The Mouse Core coordinates with the Cloning Core and Genomics Core to produce and purify the
plasmid. The Mouse Core will perform a stability assay with the plasmid and the CRISPR RNA.
Sigma supplies Mouse Core with Crispr constructs (and oligo if applicable)
Mouse Core injects Crispr construct(s) (and donor DNA if applicable)
PI genotypes toe biopsies from pups born and sends results to the Core
PI genotypes ear biopsies from pups identified as positive to confirm initial results
Mice are weaned onto PI’s smart key and protocol number and transferred to PI’s housing location.
Detailed Guide:
Before Project Begins:
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PI needs to meet with the Core Director for a free consultation to discuss the project and receive assistance in filling out the Sigma
order form.
PI is responsible for consulting with Sigma on the design of CRISPR constructs
PI is responsible for verifying screening strategy
PI is responsible for entering a project request form into PPMS: A valid smart key and protocol number must be provided.
External PI’s must submit a Purchase order for the price of the service they are requesting before the project can begin
Project Flow:
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PI will submit a project request form in PPMS
PI will submit CRISPR Order form to Sigma
Sigma will review the CRISPR order form and consult with the PI on the final design and screening strategy
When the design is finalized, Sigma will send a Quote to the Mouse Core
The Mouse Core will place the order in Emory Express for the CRISPRs and send the Purchase order to Sigma
Sigma will ship CRISPR constructs to the Mouse Core when they are completed
If project requires an oligo, Sigma will produce the oligo and ship to the Mouse Core
If project requires a plasmid,
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Sigma will send the Mouse Core the plasmid donor sequence
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The Mouse Core will forward sequence to the Cloning Core
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Cloning Core will confirm any sequence changes he requests with the PI and Sigma
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Cloning Core will produce the plasmid in cell pellet form
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Genomics Core will purify the plasmid and provide purified plasmid to the Mouse Core
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The Mouse Core Core will perform stability assay with donor DNA and CRISPR RNA to confirm DNA is not
contaminated with RNase.
The Mouse Core will perform the pronuclear injections of the CRISPR constructs (and donor DNA if applicable)
The Mouse Core will notify PI lab when pups are born.
The Mouse Core will collect toe biopsies at 7 days of age from pups born and give to PI. If PI is off campus, we will ship biopsies on
ice using PI’s FedEx account number.
PI is responsible for screening the samples and sending the results to the Mouse Core.
Once results are received by the Mouse Core and the mice are at least 14 days of age, we will collect ear biopsies from potential
founders for confirmation and give to the PI lab.
PI lab is responsible for screening the ear biopsies to confirm the genotype and sending the results to the Mouse Core
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At 4 weeks of age, the Mouse Core will wean founders onto PI’s smart key. Negative mice will be sacrificed when the positives have
been confirmed
The Mouse Core will arrange the transport of the mice to the PI’s housing location, notifying the lab of the transport.
PI lab is responsible for verifying that the lab has received the mice
Service Rate Structure:
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TMF will bill PI on the month of the project initiated:
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Sigma bills their project costs to the Mouse Core, those charges are then passed on to the PI when the injection is billed
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The Cloning Core and Genomic Core will bill their project costs to the Mouse Core, those charges are then passed on to the
PI when the injection is billed
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$5323 for Crispr Mouse Production: B6
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$5197 for Crispr Mouse Production: DB6F2 Hybrids
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$5197 for Crispr Mouse Production: FVB
Extra fee will be billed to PI lab in the following cases:
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If the Mouse Core is requested to purify DNA from tail or ear biopsies, $4 per tail
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If shipping charges are incurred transporting mice to external PIs
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If per diem fees are incurred after weaning and awaiting transport, for external PIs
Project Guarantee and Quality Control
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TMF will guarantee the existing Core protocol and Core SOP are followed