Virginia Commonwealth University

VCU Massey Cancer Center

Services and rates

The Transgenic/Knock-out Mouse Facility offers the services shown below. If you are a Massey member, the cost of these services is partially subsidized as a benefit of membership.

Transgenic / Knock-out services

Category

Description of services performed by Shared Resource staff

Generation of New GEMMs

Transgenic mouse production

  • Purify transgene
  • Microinject into fertilized mouse eggs from desired genetic background and reimplant into foster females
  • Develop genotyping strategy and screen potential founder mice
  • A minimum of 3 founders are guaranteed

Production of KO, CKO, and KI mice by ES-cell targeting

  • Electroporate ES cells with targeting vector and select clones
  • Expand and screen clones by PCR and Southern blot analysis
  • Microinject correctly targeted ES cells into blastocysts and reimplant into foster females to produce chimeras
  • Breed chimeras and screen offspring to obtain germline transmission

Production of KO, CKO, and KI mice by CRISPR/Cas9 targeting

(this has largely replaced conventional ES-cell targeting)

  • Design and synthesize sgRNA
  • Design and purchase ssODNs or construct vector encoding repair template
  • Microinject components into fertilized mouse eggs and reimplant into foster females
  • Develop genotyping strategy and screen potential founder mice
  • Breed founders and screen offspring (including sequencing) to ensure germline transmission of desired allele
  • Screen for off-target events (if requested)

GEMM Support Services

Breeding colony management

  • Acquire breeders (from JAX or other vendor, imported from other institution and rederived, or generated by TG-KO) and expand colony
  • Perform all mating, weaning, tail tissue collection, genotyping (see below), record-keeping and monitoring of colony
  • Transfer experimental animals as produced to investigator

Tail DNA genotyping

(for mice generated by TG-KO or by investigator)

  • (Investigator collects tail tissue and provides to TG-KO)
  • Perform tail DNA purification
  • Perform PCR genotyping

Mouse line rederivation by embryo transfer

  • Facilitate imports in conjunction with the Division of Animal Resources (DAR) as needed (imported breeders are placed in designated "rederivation room")
  • Expand colony if needed; purchase female breeders if needed
  • Superovulate females and mate to males
  • Collect fertilized eggs, transfer to barrier facility, and reimplant into foster females
  • Screen offspring for desired allele(s) (if requested)
  • Transfer of rederived mice to investigator

Embryo cryopreservation

  • Obtain male breeders from investigator
  • Purchase females or obtain from investigator
  • Superovulate females and mate to males
  • Collect fertilized eggs and transfer to barrier facility
  • Perform cryopreservation and provide long-term storage in LN2
  • A minimum of 200 embryos are cryopreserved

Sperm cryopreservation

  • Collect sperm from 2 male mice obtained from investigator
  • Perform cryopreservation using JAX kit
  • Ship straws to JAX for QC and long-term storage

Management of IVIS spectrum imager in MMRB vivarium

  • Provide initial training to new users
  • Oversee sign-up for use through CORES
  • Perform routine maintenance of facility; maintain O2 supply
  • Schedule annual maintenance and renew service contract

Consulting/Training

For all projects:

  • Meetings to discuss experimental objectives, details of experimental design, optimal/alternative strategies, etc

Other examples:

  • Breeding and genotyping strategy consultation; genotyping trouble-shooting
  • Basic training on use of IVIS imager
  • Determining whether desired mouse lines are available from JAX or various consortia or repositories
  • Facilitating import of lines from other institutions (in conjunction with DAR)
  • Provision of support letters for grant applications (18 letters provided in 2015)
  • Assisting with budget preparation for grants including GEMM production and breeding
  • Assisting with preparation of IACUC protocols

 

Technologies provided

Instrument

Description

IVIS Spectrum Optical Imager

  • The TG-KO oversees management of the IVIS optical imager in the MMRB vivarium and provides end-user training

 

Rates and subsidy

Note that rates are current as of August 1, 2016, but are subject to change. Massey members pay 75 percent of these rates per the member subsidy program.

Service

Price

Generation of New GEMMs

Transgenic mouse production

$4,500

Production of KO, CKO and KI mouse lines by ES-cell targeting

  • ES-cell electroporation, selection and clone expansion

$5,500

  • ES-cell screening by PCR and Southern blot – first two 96-well plates
  • ES-cell screening by PCR and Southern blot – each additional plate

$2,000

$1,000

  • Blastocyst injection for chimera production

$3,500

  • Breeding to obtain germline transmission

See colony management

Production of KO and KI mouse lines by CRISPR/Cas9

  • Reagent design, synthesis, purchase

$2,500

  • Microinjection and screening

$4,500

Production of CKO (floxed) mouse lines by CRISPR/Cas9

  • Reagent design, synthesis, purchase

$3,500

  • Microinjection and screening

$6,500

GEMM Support Services

Breeding colony management

$40/hour

Genotyping – tail DNA prep

Genotyping – PCR (per reaction)

$5

$5

Mouse line rederivation by embryo transfer

$1,000

Embryo cryopreservation

$2,500 plus cost of mice purchased

Sperm cryopreservation (includes QC and 3 years’ storage at JAX)

$1,200

Mouse line cryorecovery

$1,000

Use of IVIS imager

$25/hour

Consulting/Training

All consultation

No charge

Massey member subsidy program

As of January 1, 2012, VCU Massey subsidizes 25 percent of the cost of members’ core laboratory usage for the following Massey-supported, fee-for-service shared resources:

  • Biological Macromolecule Shared Resource Core
  • Flow Cytometry Shared Resource Core
  • Structural Biology Shared Resource Core
  • Tissue and Data Acquisition and Analysis Core
  • Transgenic/Knock-out Mouse Shared Resource Core

Note that this subsidy is available only for Massey members. To receive this member benefit, please initiate core lab services using a VCU index code associated with your name. When you are billed for core laboratory usage, the CORES system will automatically split the invoice between your index code and a Massey account established for this purpose. You will be charged 75 percent of the published rate and VCU Massey will provide a 25 percent co-pay to cover the balance.

Please direct any questions or concerns about the member subsidy benefit to Paul Fawcett, Ph.D. at pfawcett@vcu.edu or (804) 833-1959.

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