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Generation of a germ cell‐specific mouse transgenic cherry reporter, Sohlh1‐mCherryFlag
Author(s) -
Suzuki Hitomi,
Dann Christina Tenenhaus,
Rajkovic Aleksandar
Publication year - 2013
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.22347
Subject(s) - biology , germ cell , transgene , microbiology and biotechnology , spermatogenesis , germline , oogenesis , genetically modified mouse , stem cell , ovary , gene , genetics , oocyte , endocrinology , embryo
Visualization of differentiating germ cells is critical to understanding the formation of primordial follicles in the ovary, and the commitment of spermatogonial stem cells to differentiation. We engineered and generated a BAC transgenic mouse line, Sohlh1‐mCherryFlag ( S1CF ), under the direction of the native Sohlh1 promoter. Sohlh1 is a germ cell‐specific gene that encodes the basic helix‐loop‐helix (bHLH) transcriptional regulator that is essential in oogenesis and spermatogenesis. Sohlh1 expression is unique, and is limited to perinatal and early follicle oocytes and differentiating spermatogonia. The Sohlh1‐mCherryFlag transgene was engineered to fuse SOHLH1 to the red fluorescent protein CHERRY with 3‐tandem‐FLAG tags. S1CF animals fluoresce specifically in the oocytes of perinatal ovaries and small follicles in adult ovaries, as well as in spermatogonia, a pattern that is similar to endogenous SOHLH1. Moreover, S1CF rescued germ cell loss and infertility in both male and female Sohlh1 −/− animals. The FLAG‐tag on S1CF was effective for immunostaining and immunoprecipitation. The Sohlh1‐mCherryFlag transgenic mouse provides a unique model to study early germ cell differentiation, as well as in vivo imaging and purification of differentiating germ cells. genesis 51:50–58, 2013. © 2012 Wiley Periodicals, Inc.