PUBLICATION

Chemical Genetics: Manipulating the Germline with Small Molecules

Authors
Jin, Y.N., Peterson, R.T.
ID
ZDB-PUB-210220-28
Date
2021
Source
Methods in molecular biology (Clifton, N.J.)   2218: 61-73 (Chapter)
Registered Authors
Peterson, Randall
Keywords
3′UTR, Chemical genetics, Ddx4/Vasa, Embryo, Germline, Germline stem cells, Nanos3, Primordial germ cells, Small-molecule screens, Zebrafish
MeSH Terms
  • 3' Untranslated Regions/genetics
  • Animals
  • DEAD-box RNA Helicases/genetics
  • Embryo, Nonmammalian/drug effects
  • Embryonic Development/drug effects
  • Embryonic Development/genetics
  • Female
  • Germ Cells/drug effects*
  • Luminescent Proteins/genetics
  • Male
  • RNA-Binding Proteins/genetics
  • Small Molecule Libraries/pharmacology*
  • Zebrafish/genetics
  • Zebrafish Proteins/genetics
PubMed
33606223 Full text @ Meth. Mol. Biol.
Abstract
Primordial germ cells (PGCs) are the precursor cells that form during early embryogenesis and later differentiate into oocytes or spermatozoa. Abnormal development of PGCs is frequently a causative factor of infertility and germ cell tumors. However, our understanding of PGC development remains insufficient, and we have few pharmacological tools for manipulating PGC development for biological study or therapy. The zebrafish (Danio rerio) embryos provide an excellent in vivo animal model to study PGCs, because zebrafish embryos are transparent and develop outside the mother. Importantly, the model is also amenable to facile chemical manipulations, including scalable screening to discover novel compounds that alter PGC development. This chapter describes methodologies for manipulating the germline (i.e., PGCs) with small molecules and for monitoring PGC development. Utilizing the 3'UTR of PGC marker genes such as nanos3 and ddx4/vasa is a key component of these methodologies, which consist of expressing fluorescent or luminescent proteins in PGCs, treatment with small molecules, and quantitative observation of PGC development.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping