| RRC ID |
87148
|
| 著者 |
Le HT, Hasegawa Y, Daitoku Y, Kato K, Miznuo-Iijima S, Dinh TTH, Kuba Y, Osawa Y, Mikami N, Morimoto K, Ayabe S, Tanimoto Y, Murata K, Yagami KI, Takahashi S, Mizuno S, Sugiyama F.
|
| タイトル |
Generation of B6-Ddx4em1(CreERT2)Utr , a novel CreERT2 knock-in line, for germ cell lineage by CRISPR/Cas9.
|
| ジャーナル |
Genesis
|
| Abstract |
Germ cell development is essential for maintaining reproduction in animals. In postpubertal females, oogenesis is a highly complicated event for producing fertilizable oocytes. It starts when dormant primordial oocytes undergo activation to become growing oocytes. In postpubertal males, spermatogenesis is a differentiation process for producing sperm from spermatogonial stem cells. To obtain full understanding of the molecular mechanisms underlying germ cell development, the Cre/loxP system has been widely applied for conditional knock-out mouse studies. In this study, we established a novel knock-in mouse line, B6-Ddx4 em1(CreERT2)Utr , which expresses CreERT2 recombinase under the control of the endogenous DEAD-box helicase 4 (Ddx4) gene promoter. Ddx4 was specifically expressed in both female and male germ cell lineages. We mated the CreERT2 mice with R26GRR mice, expressing enhanced green fluorescent protein (EGFP) and tDsRed before and after Cre recombination. We found tDsRed signals in the testes and ovaries of tamoxifen-treated B6-Ddx4 em1(CreERT2)Utr ::R26GRR mice, but not in untreated mice. Immunostaining of their ovaries clearly showed that Cre recombination occurred in all oocytes at every follicle stage. We also found 100% Cre recombination efficiency in male germ cells via the progeny test. In summary, our results indicate that B6-Ddx4 em1(CreERT2)Utr is beneficial for studying female and male germ cell development.
|
| 巻・号 |
58(7)
|
| ページ |
e23367
|
| 公開日 |
2020-7-1
|
| DOI |
10.1002/dvg.23367
|
| PMID |
32293787
|
| MeSH |
Animals
Cell Lineage*
DEAD-box RNA Helicases / genetics*
DEAD-box RNA Helicases / metabolism
Female
Gene Knock-In Techniques / methods*
Germ Cells / cytology
Germ Cells / metabolism*
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Integrases / genetics*
Integrases / metabolism
Male
Mice
Mice, Inbred C57BL
Promoter Regions, Genetic
|
| IF |
1.76
|
| リソース情報 |
| 遺伝子材料 |
pBs-gsg P2A-Cre ERT2 (RDB14416) |
