Overview
The GMF provides a comprehensive-service option for the generation of mice using ES cells or targeting vectors from the International Knockout Mouse Consortium (IKMC). This consortium comprises all major large-scale mutagenesis centers such as KOMP, EUCOMM, NorComm and TIGM.
To get started, submit a GMF Service Request Form indicating what type of reagents, genetic locus, IKMC project, as well as ID of ES cell clones (if possible, we recommend ordering 2 to 3 clones).
For each project, project investigator handles the following tasks:
1. Submitting orders to the respective IKMC center
2. Filing MTA with the institution, and submitting payments
3. Receiving ES cell clones or targeting vectors, and then deliver them to GMF. Or, you request the materials to be shipped directly to GMF: Genome Modification Facility, Harvard University, BioLab # 1062, 16 Divinity Avenue, Cambridge, MA 02138
The GMF handles the following tasks:
4. Producing chimeric mice or transfected ES cell clones
a) for generating chimeric mice, we routinely use albino-B6 host embryos for ES cells, JM8N4 (black) JM8A3.N1 or JM8A1.N3 (agouti). For details, check ES cell Blastocyst Injection service b) with targeting vectors, we use either the JM8A3.N1 (B6N) cell line from KOMP-CSD/EUCOMM or V6.5 (129xB6F1), depending on request. For details, check our ES Cell Gene Targeting service5. Transferring chimeric mice to investigator's colony
Background on IKMC
The International Knockout Mouse Consortium, IKMC, consists of several independent centers, including KOMP-CSD, KOMP-Regeneron/Velocigene, EUCOMM, NorCOMM, and TIGM, each having specific objectives regarding the number and type of targeted loci in ES cells they produce. An overview of the specific goals of the respective centers can be found on the IKMC homepage. Notably, some of the centers, such as KOMP-CSD, EUCOMM, and NorCOMM, generate vectors and ES cell clones that have CONDITIONAL POTENTIAL. The others, including KOMP-Regeneron and TIGM, produce trapped or targeted loci that give rise to straight germ line gene knockouts. They all can be accessed through the IKMC website.
GMF Track Record using KOMP and EUCOMM ES Cell Clones
The table below summarizes mouse production in the GMF using KOMP/EUCOMM reagents. By the end of January 2012, the GMF had processed 33 orders. The genetic loci, source of ES cell clones or vectors, and status are shown.
Shown below are examples of chimeras produced using JM8 and E14 ES cell clones most of which injected into albino-B6 host embryos. Details are described in the pictures.
Estimated Time Frame and Costs for Mouse Production using KOMP/EUCOMM Reagents
|
Steps |
Action Items |
Estimated Time Frame (during week #) |
Fees |
|
1 |
Submit Service Request Form |
1 |
TBD |
|
2 |
GMF processes Request and Assigns a Job ID |
1 |
TBD |
|
3 |
Submit and finalize MTA with OTD |
1 |
TBD |
|
4 |
Generate PO# for UC Davis (KOMP) or Helmholtz Center (EUCOMM) |
TBD | |
|
5 |
Submit Payment |
2 |
TBD $ amount on invoice from UC Davis or Helmholz Center |
|
6 |
UC Davis (KOMP) or Helmholtz Center (EUCOMM) initiate validation of reagents |
12 (KOMP) 16-20 (EUCOMM) |
TBD |
|
Production of chimeras in GMF |
14-22 |
TBD |
|
|
8 |
Transfer chimeras to PI colony |
20-28 |
TBD |
|
9 |
Service invoiced |
20-28 |
Total, plus per diem costs |
Total Estimated Time: 20-28 Weeks
Disclaimer The Genome Modification Facility at Harvard University does not guarantee that germ line transmitting chimeras will be obtained from ES cells procured from any of the IKMC centers. Non-Harvard Academic & Non-Profit users pay 15% more than Harvard users. Service fees for Industry & For-Profit users are given upon request. The GMF further retains the rights to change fees without prior notice.(Return to Services and Fees listing)