A novel method to identify syncytiotrophoblast-derived RNA products representative of trisomy 21 placental RNA in maternal plasma

Methods Mol Biol. 2008:444:291-302. doi: 10.1007/978-1-59745-066-9_23.

Abstract

A novel in vitro method is described wherein gene expression profiling is reflective and informative for the way how syncytiotrophoblast cells shed RNA products in vivo in maternal plasma. After controlled denudation, RNA is obtained selectively from the syncytiotrophoblast cells of trisomy 21 placentae. cDNA copies are subsequently analyzed by microarray profiling and cDNA cloning with sequencing. Given the preponderance of 5' mRNA fragments lacking a poly-A tail, the placental RNA products are amplified after polymerase A-mediated tailing by using a method originally designed for small-sized microRNAs. This approach, when combined with cDNA library or microarray expression screening, is a novel in vitro method to screen for syncytiotrophoblast-derived RNA products representative of trisomy 21 placental RNA as present in vivo in maternal plasma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • DNA Mutational Analysis
  • Down Syndrome / diagnosis*
  • Down Syndrome / genetics
  • Female
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental
  • Genetic Testing*
  • Humans
  • Maternal-Fetal Exchange
  • Oligonucleotide Array Sequence Analysis
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • RNA, Messenger / blood*
  • RNA, Messenger / isolation & purification
  • Trophoblasts / metabolism*

Substances

  • RNA, Messenger