Towards Functional Annotation of the Preimplantation Transcriptome: An RNAi Screen in Mammalian Embryos

Sci Rep. 2016 Nov 21:6:37396. doi: 10.1038/srep37396.

Abstract

With readily available transcriptome-wide data, understanding the role of each expressed gene is an essential next step. Although RNAi technologies allow for genome-wide screens in cell culture, these approaches cannot replace strategies for discovery in the embryo. Here we present, for the first time, a knockdown screen in mouse preimplantation embryos. Early mammalian development encompasses dynamic cellular, molecular and epigenetic events that are largely conserved from mouse to man. We assayed 712 genes for requirements during preimplantation. We identified 59 genes required for successful development or outgrowth and implantation. We have characterized each phenotype and revealed cellular, molecular, and lineage specific defects following knockdown of transcript. Induced network analyses demonstrate this as a valid approach to identify networks of genes that play important roles during preimplantation. Our approach provides a robust and efficient strategy towards identification of novel phenotypes during mouse preimplantation and facilitates functional annotation of the mammalian transcriptome.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blastocyst / metabolism*
  • Embryo, Mammalian / cytology
  • Embryo, Mammalian / metabolism*
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques
  • Microinjections
  • Molecular Sequence Annotation*
  • Morula / metabolism
  • Phenotype
  • RNA Interference*
  • RNA, Double-Stranded / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Transcriptome / genetics*

Substances

  • RNA, Double-Stranded
  • RNA, Messenger