The generation and analysis of genetically modified mice, including transgenic and knockout animals, is an essential technology for cancer research. The Transgenic Mouse Shared Resource makes these powerful technologies available to all Cancer Center members.
- Production of transgenic mice by pronuclear DNA microinjection and embryo transfer.
- 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 mice.
- CRISPR/Cas9 gene editing in mouse embryos
- 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 Genetics Resource
CRISPR/Cas9 Genome Editing Service
TMSR 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.
- You may read this article to get general information about CRISPR/Cas9 genome editing in mouse embryos. For special interest in HDR with CRISPR, please read this article and this article. It is reported in this article that a guide ends in GG motif (not the NGG in PAM sequence) tends to be a more efficient guide. This article is the most recent reference for designing a better guide RNA and the server is CRISPRscan.
- TMSR 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. TMSR 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
TMSR 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. TMSR 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).
- TMSR provides PMSG and hCG for superovulation of these female blastocyst donors.
- Inject 100ul PMSG (available from TMSR) IP into each mouse at 4pm on Tuesday.
- Inject 100ul hCG (available from TMSR) IP into each mouse at 3pm 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 11am next Monday.