| RRC ID |
57970
|
| 著者 |
Serizawa T, Isotani A, Matsumura T, Nakanishi K, Nonaka S, Shibata S, Ikawa M, Okano H.
|
| タイトル |
Developmental analyses of mouse embryos and adults using a non-overlapping tracing system for all three germ layers.
|
| ジャーナル |
Development
|
| Abstract |
Genetic lineage-tracing techniques are powerful tools for studying specific cell populations in development and pathogenesis. Previous techniques have mainly involved systems for tracing a single gene, which are limited in their ability to facilitate direct comparisons of the contributions of different cell lineages. We have developed a new combinatorial system for tracing all three germ layers using self-cleaving 2A peptides and multiple site-specific recombinases (SSRs). In the resulting TRiCK (TRiple Coloured germ layer Knock-in) mice, the three germ layers are conditionally and simultaneously labelled with distinct fluorescent proteins via embryogenesis. We show that previously reported ectopic expressions of lineage markers are the outcome of secondary gene expression. The results presented here also indicate that the commitment of caudal axial stem cells to neural or mesodermal fate proceeds without lineage fluctuations, contrary to the notion of their bi-potency. Moreover, we developed IMES, an optimized tissue clearing method that is highly compatible with a variety of fluorescent proteins and immunostaining, and the combined use of TRiCK mice and IMES can facilitate comprehensive analyses of dynamic contributions of all three germ layers.
|
| 巻・号 |
146(21)
|
| 公開日 |
2019-11-4
|
| DOI |
10.1242/dev.174938
|
| PII |
dev.174938
|
| PMID |
31597657
|
| MeSH |
Animals
Brain / metabolism
Cell Lineage*
Crosses, Genetic
DNA Nucleotidyltransferases / metabolism
Embryonic Stem Cells / cytology
Endoderm / cytology
Endothelium, Vascular / cytology
Female
Gene Expression Regulation, Developmental*
Gene Knock-In Techniques*
Genotype
Germ Layers / cytology*
Green Fluorescent Proteins / metabolism
HEK293 Cells
Heart / embryology
Humans
Imaging, Three-Dimensional
Liver / embryology
Male
Mesoderm / cytology
Mice
Mice, Inbred C57BL
Myocardium / cytology
Neural Plate / cytology
|
| IF |
5.611
|
| 引用数 |
0
|
| リソース情報 |
| 遺伝子材料 |
pCAGGS-Cre (RDB08998)
B6N Mouse BAC clone (RDB07573)
B6Ng01-316F09, B6Ng01-307M06, B6Ng01-307A03, and B6Ng01-211A18 |
| ヒト・動物細胞 |
B6NJ-22^(UTR)(AES0141) |
