RRC ID 57970
Author Serizawa T, Isotani A, Matsumura T, Nakanishi K, Nonaka S, Shibata S, Ikawa M, Okano H.
Title Developmental analyses of mouse embryos and adults using a non-overlapping tracing system for all three germ layers.
Journal 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.
Volume 146(21)
Published 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
Times Cited 0
DNA material pCAGGS-Cre (RDB08998) B6N Mouse BAC clone (RDB07573) B6Ng01-316F09, B6Ng01-307M06, B6Ng01-307A03, and B6Ng01-211A18
Human and Animal Cells B6NJ-22^(UTR)(AES0141)