Successful in vitro fertilization and generation of transgenics in Black and Tan Brachyury (BTBR) mice.

Transgenic Res

Division of Endocrinology, Diabetes and Metabolism, Department of Medicine, University of Wisconsin-Madison, 4147 MFCB, 1685 Highland Ave, Madison, WI, 53705, USA.

Published: December 2016

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 backcrossing, 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.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5106318PMC
http://dx.doi.org/10.1007/s11248-016-9974-0DOI Listing

Publication Analysis

Top Keywords

btbr
10
vitro fertilization
8
black tan
8
tan brachyury
8
brachyury btbr
8
btbr mice
8
btbr mouse
8
success btbr
8
embryo manipulations
8
successful vitro
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!