Measurement of mRNA Poly(A) Tail Lengths in Drosophila Female Germ Cells and Germ-Line Stem Cells

Methods Mol Biol. 2017:1463:93-102. doi: 10.1007/978-1-4939-4017-2_7.

Abstract

mRNA regulation by poly(A) tail length variations plays an important role in many developmental processes. Recent advances have shown that, in particular, deadenylation (the shortening of mRNA poly(A) tails) is essential for germ-line stem cell biology in the Drosophila ovary. Therefore, a rapid and accurate method to analyze poly(A) tail lengths of specific mRNAs in this tissue is valuable. Several methods of poly(A) test (PAT) assays have been reported to measure mRNA poly(A) tail lengths in vivo. Here, we describe two of these methods (PAT and ePAT) that we have adapted for Drosophila ovarian germ cells and germ-line stem cells.

Keywords: Deadenylation; Drosophila; Germ line; Germ-line stem cell; Ovary; PAT assay; Poly(A) tail; Polyadenylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Drosophila / genetics*
  • Female
  • Gene Expression Regulation
  • Ovary / chemistry*
  • Ovary / cytology
  • Poly A / analysis*
  • Polyadenylation
  • RNA, Messenger / chemistry*
  • Stem Cell Niche
  • Stem Cells / chemistry
  • Stem Cells / cytology

Substances

  • RNA, Messenger
  • Poly A