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Services and rates

Massey members receive a 25 percent subsidy for the cost of these services.

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 designatedrederivation 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. 

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

Massey subsidizes 25 percent of the cost a member's 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

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.