Characterization of a Dazl-GFP germ cell-specific reporter.
In this study, we characterized the promoter activity of a 1.7 kb sequence in the 5' flanking region of the mouse Deleted in Azoospermia-Like (Dazl) gene. We found the 1.7 kb sequence sufficient to drive robust germ cell-specific expression of green fluorescent protein (GFP) in adult mouse testis and lower levels of expression in adult ovary and in fetal and newborn gonads of both sexes. This expression pattern was confirmed in two independently-derived transgenic mouse lines. In adult testis, Dazl-GFP exhibited a developmentally-regulated, stage-specific expression pattern during spermatogenesis. GFP was highly expressed in spermatocyte stages, with strongest expression in pachytene spermatocytes. Weaker expression was observed in round and elongating spermatids, as well as spermatogonial cells. In the fetal gonad, GFP transcript was detected by e12.5 in both sexes; however, GFP fluorescence was only detected during later embryonic stages. In addition, we produced mouse embryonic stem cell (ESC) lines harboring the Dazl-GFP reporter and used this reporter to isolate putative germ cell populations derived from mouse ESCs following embryoid body differentiation and fluorescence activated cell sorting.
Nicholas
Cory R
CR
Department of Obstetrics and Gynecology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Palo Alto, CA, USA. cory2007@stanford.edu
Xu
Eugene Y
EY
Banani
Salman F
SF
Hammer
Robert E
RE
Hamra
F Kent
FK
Reijo Pera
Renee A
RA
D020121:5' Untranslated Regions; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D002454:Cell Differentiation; D002469:Cell Separation; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009866:Oogenesis / Q000235:genetics; D010053:Ovary / Q000166:cytology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism*; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D016601:RNA-Binding Proteins / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D013091:Spermatogenesis / Q000235:genetics; D013737:Testis / Q000166:cytology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism*
D020121:5' Untranslated Regions; C508515:DAZL protein, mouse; D017931:DNA Primers; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins