Genetically Modified Mouse Models

The generation and analysis of genetically modified mice, including transgenic and knockout animals, is an essential technology for cancer research. The Genetically Modified Mouse Models Shared Resource (GMMMSR) makes these powerful technologies available to all Cancer Center members.

Services include:

• Production of transgenic mice by pronuclear DNA microinjection and embryo transfer
• CRISPR/Cas9 gene editing in mouse embryos
• Production of chimeric mice by blastocyst injection of genetically modified ES cells and embryo transfer, which is part of the procedure of producing knockout or knockin mic
• Gene targeting, which involves the electroporation of a targeting vector into pluripotent mouse embryonic stem (ES) cells and the selection of gene-targeted clones.
• Education of investigators and advice on vector and experimental design for experiments using transgenic and knockout mice.
• Mouse genetic resources

GMMMSR’s Full Gene Targeting Service and Terms

GMMMSR offers a comprehensive gene targeting service in 129S6/SvEvTac, 129B6 hybrid, and C57BL/6N ES cells. Clients provide the details of the genetic modification to be introduced into the mouse genome. GMMMSR will design the gene targeting vector, target the ES cells with the intended genetic modification, generate chimeric mice using targeted ES cells, and perform a germline transmission test, all within a 12-month timeframe.

Service Details:

• Duration: This service period lasts for 12 months from the completion of an iLab service request.
• Agreement: Before beginning the full gene targeting project, GMMMSR and the client must agree on the DNA sequence (genetic modification) to be introduced into the mouse genome. GMMMSR guarantees the delivery of heterozygous mice with the agreed-upon genetic modification within 12 months. However, GMMMSR cannot guarantee the functionality of the modification, such as the expression of the reporter or the functional switch of the introduced genetic switch.
• Termination: Either GMMMSR or the client may terminate this full gene targeting service after the 12-month service period. If either party decides to terminate the service, GMMMSR will refund 100% of the service charge to the client. If the client terminates the project before the 12-month service period ends, GMMMSR will deduct the cost of any reagents and labor already used during the project and refund the client's account accordingly.

CRISPR/Cas9 Genome Editing Service

GMMMSR provides CRISPR genome editing service in B6CBAF1 or C57BL/6 backgrounds with either CRISPR RNAs or plasmids. Please contact Victor Lin email: CSL5@CUMC.COLUMBIA.EDU or call 1-212-851-4716 for details.

• For general information about CRISPR/Cas9 genome editing in mouse embryos, please refer to this NCBI article, "A Mouse Geneticist’s Practical Guide to CRISPR Applications".
• For special interest in HDR with CRISPR, please refer to
  • "Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining" 
  • "Cloning-free CRISPR/Cas system facilitates functional cassette knock-in in mice"
• "Dramatic Enhancement of Genome Editing by CRISPR/Cas9 Through Improved Guide RNA Design" reports that a guide ends in GG motif (not the NGG in PAM sequence) tends to be a more efficient guide. 
• "CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo." This article is the most recent reference for designing a better guide RNA and the server is CRISPRscan.

GMMMSR guarantees 10 pups or two founders (whichever comes first) for each CRISPR genome editing service request. Genome editing is not guaranteed as it is dependent on the quality of sgRNAs. GMMMSR suggests that 3-4 sgRNAs should be tested in vitro for each project to identify the guide(s) that doesn't work in vitro and test the guides that work in vitro in mouse embryos.

ES Cell Derivation Service

GMMMSR now offers mouse embryonic stem (ES) derivation service. It has become increasingly popular to derive ES cells from genetically modified mice carrying specific genetic modification and use these ES cells for research.
Mouse ES cells are routinely derived from C57BL/6 and 129 mouse strains. GMMMSR suggests our clients breed their mice into these two backgrounds in order to achieve higher ES cell derivation success rate.

What clients should do:

• Clients should breed 5 female (9-13 grams body weight, about three weeks old) blastocyst donors and 5 stud males (2-3 months old). These mice should be pathogen-free (We will check the health status with ICM before we schedule an ES cell derivation service).
• GMMMSR provides PMSG and hCG for superovulation of these female blastocyst donors.
• Inject 100ul PMSG (available from GMMMSR) IP into each mouse at 4 p.m. on Tuesday.
• Inject 100ul hCG (available from GMMMSR) IP into each mouse at 3 p.m. on Thursday. Pair each PMSG/hCG-primed female with one stud male right after hCG injection.
• Pick out the females and place these five females in one cage Friday morning
• Bring these five female mice to ICRC604 at 11 a.m. next Monday.

Locations and Contacts

The Genetically Modified Mouse Models Models shared resource is located at:

Irving Cancer Research Center
1130 St. Nicholas Avenue, Room 604
Tel: 212-851-4717 Fax: 212-851-4715

Contacts Chyuan-Sheng (Victor) Lin, PhD, Manager
Tel: 212-851-4716
E-mail: CSL5@columbia.edu

User Fees and Policies

The following policies are in place for the Genetically Modified Mouse Models Models shared resource relating to scheduling services and charges for use of the facility.

Service Request

Investigators who would like to request services from the facility should log in to iLab at https://cumc.corefacilities.org with their UNI and password, select Transgenic Mouse from the list of cores, and open the Request Services tab. Follow the instructions to initiate a request and fill in the appropriate forms. Contact the TNSR at cls5@cumc.columbia.edu if additional help is needed in making a request. Our services are provided based on the order in which the request is received.

User Fees

Notes On Services

• Thawing of frozen ES clones for microinjection will incur a charge for each clone thawed at a rate of $100 (CC Members) or $125 per clone (Non-Members).
• After the first clone of ES Cells, each additional clone for the same experiment will be $100 (CC Members) or $125 (Non-Members).
• GMMMSR will expand each ES line to the well of 24-well plate and freeze two vials for each ES line. 50ul ES lysate will be provided for genotyping purpose. Setup charge will be waived if no ES line is obtained.

Other Policies

• DNA samples and ES cell lines are injected in the order in which the signed service request forms are received. Currently, the average time from receipt of the service request to injection is approximately 2 weeks.
• For ES Cell electroporation and expansion, success in gene targeting cannot be guaranteed due to inherent variability in the frequency of targeting different loci, and the possibility of mistakes or mutations in the targeting vector.

Online Reservations

The Biomedical Informatics Shared Resource uses the iLab Core Management System for service requests and billing. Before you can request services, you must log into iLab and be approved as a lab member.