Transgenic Core

Director: Arlene Sharpe

 
 
Core Summary:

The BWH Transgenic Core, established in 1992 by Dr. Arlene Sharpe, provides gene targeting and microinjection services using state-of the ar facilites and equipment.

The Core has extensive experience generating transgenic mice using DNA constructs (including BAC DNAs) and lentiviral constructs. We also have a track record of success generating KO/KI mice by performing gene targeting, or by expanding targeted ES Cells from consortia. And in recent years, we have expanded our services to include CRISPR injections and cryopreservation.

The Core works with investigators to achieve optimal outcomes for each project.

Personnel/Contact Information:

Director: Arlene Sharpe, M.D, Ph.D.
Phone: (617) 432-6569
Fax: (617) 432 -6570
Email: arlene_sharpe@hms.harvard.edu

Supporting Staff:

Transgenic Specialist: Lina Du, M.D.
Phone: (617) 432-6227
Email: lina_du@hms.harvard.edu

ES Cell Culture Technician: Michael Peluso
Phone: (617) 432-6182
Email: bwhtransgeniccore@gmail.com

 
Services:

    Click for New Services Flyer

  • Microinjection of DNA constructs (including BACs) into pronuclei for generating transgenic mice
    (FVB or C57 BL/6 strain background)
  • Microinjection of embryonic stem (ES) cells into blastocysts for generating knockout mice
  • ES cell transfection for generating ES cell clones with the targeted combination event
  • Lentiviral transgenics
  • Microinjection of IPS
  •  

FY15 User Fees (Effective 10/01/14) :

Service
Internal Users
External Users
Partners
Non-Partners AMC & Non-Profit
For-Profit & Industry
Lentiviral Injection
$5,010
$7,215
$7,966
DNA (FVB) Pronuclear Injection
$4,312
$6,210
$6,857
DNA (C57) Pronuclear Injection
$4,264
$6,141
$6,780
DNA Injection & Microdissection
$2,160
$3,111
$3,435
Additional DNA Injection
$2,351
$3,386
$3,739
BAC Injection
$4,887
$7,038
$7,771
ES Cell Injection (129)
$2,360
$3,399
$3,753
ES Cell Injection (B6)
$2,439
$3,513
$3,879
Tail Biopsy
$175
$252
$279
Electroporation (129) ES
$2,877
$4,143
$4,575
Electroporation (B6) ES
$3,352
$4,827
$5,330
ES Expansion
$700
$1,008
$1,113
ES Cell Prep for Injection
$275
$396
$438
Balb C
$5,325
$7,668
$8,467
IPS
$2,935
$4,227
$4,667
CRISPR Injection
$2,345
$3,377
$3,729
Sperm Cryopreservation
$400
$576
$636
Embryo Cryopreservation
$550
$792
$875
Thawing and Implantation of Embryos
$550
$792
$875
Fresh Sperm IVF
$1,400
$2,016
$2,226
Frozen Sperm IVF
$1,400
$2,016
$2,226
Exportation Fee
$35.00
$50.40
$55.65
Technical Services
$75
$108
$120
Animal Per Diem
$75
$108
$120

 

Outline & Timeline of Project:

Our ES facility will generate targeted ES cell lines with KO/KI constructs provided by the investigator.
Upon receipt of the construct and application form, an electroporation date will be scheduled. The Core
Director, Dr. Arlene Sharpe, will prioritize the service requests according to the difficulty of the project
and workload at the time of receipt.

1. Transfection of your purified linearized DNA construct into ES cells, JM8 (C57 BL/6), J1 (129/SV)
and Bruce 4 (C57 BL/6J).
2. After the electroporation and selection, clones will be picked (192), cultured in multi-well plates
and replicated. The clones will then be frozen in order to expand the positive clones after
screening. This part of the process usually takes 3 weeks to complete.
3. The investigator will be notified and given DNA isolated from each clone suitable for screening by
Southern and PCR.
4. Since storage of frozen cells is not optimal, it is strongly suggested that investigators return the
screening results to the core within 3 months of receipt of the DNA samples.
5. After receiving screening results, up to 12 ES positive clones will be expanded, split for DNA, and
frozen down. The investigator will then receive DNA from each clone for reconfirmation of
targeted recombination. This process usually takes 2 weeks.
6. After reconfirmation, the best clone will then be scheduled for injection into blastocysts. Chimeric
mice will be given to the investigator.

Please visit the Partners Core Management System to Request Services:
https://researchcores.partners.org/login

Click Here for Instructions on How to Request Services

 

 

 
 
     
 

 

Cores Home Page
Directory of Cores
Forms, Tools and Resources