期刊
TRANSGENIC RESEARCH
卷 25, 期 6, 页码 847-854出版社
SPRINGER
DOI: 10.1007/s11248-016-9974-0
关键词
BTBR; In vitro fertilization; Embryo transfer; Transgenic mouse model; Diabetes mellitus; Autism
类别
资金
- National Institutes of Health [T32 RR023916, T32 OD010423]
- National Institute of Aging [T32 AG000213]
- University of Wisconsin-Madison Science and Medicine Graduate Research Scholars Program
- NIDDK [DK083442]
- University of Wisconsin Department of Medicine
The Black and Tan Brachyury (BTBR) mouse strain is a valuable model for the study of long-term complications from obesity-induced type 2 diabetes mellitus and autism spectrum disorder. Due to technical difficulties with assisted reproduction, genetically modified animals on this background have previously been generated through extensive back-crossing, which is expensive and time-consuming. We successfully generated two separate transgenic mouse lines after direct zygote microinjection into this background strain. Additionally, we developed in vitro fertilization (IVF) methods for the BTBR mouse. We found low rates of fertilization and implantation in this strain, and identified the BTBR oocyte as the primary culprit of low success with BTBR IVF. We achieved an increase in live born pups from 5.9 to 35.6% with IVF in the BTBR strain by use of BTBR females at a younger age (18-25 days), collection of oocytes 15-17 h after superovulation, and the use of supplemented fertilization media. This method eliminates the need for time consuming assisted embryo manipulations that are otherwise required for success with BTBR oocytes. This advancement provides an exciting opportunity to directly generate BTBR transgenics and gene-edited mice using both traditional and emerging genomic editing techniques, such as CRISPR/Cas9. These methods also allow effective colony preservation and rederivation with these strains. To our knowledge, this is the first report describing embryo manipulations in BTBR mice.
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