Transfection of Small Noncoding RNAs into Arabidopsis thaliana Protoplasts

Methods Mol Biol. 2020:2166:413-429. doi: 10.1007/978-1-0716-0712-1_24.

Abstract

Polyethylene glycol transfection of plant protoplasts represents an efficient method to incorporate foreign DNA and study transient gene expression. Here, we describe an optimized protocol to deliver small noncoding RNAs into Arabidopsis thaliana protoplasts. An example of application is provided by demonstrating the incorporation of a 20 nt long small noncoding RNA deriving from the 5' extremity of an A. thaliana cytosolic alanine tRNA into freshly isolated protoplasts.

Keywords: Northern blot; Protoplasts; Small noncoding RNAs; Transfection; tRNA-derived fragments.

MeSH terms

  • Alanine / genetics
  • Arabidopsis / genetics*
  • Protoplasts / metabolism*
  • RNA, Plant / genetics*
  • RNA, Small Untranslated / genetics*
  • RNA, Transfer / genetics
  • Transfection / methods*

Substances

  • RNA, Plant
  • RNA, Small Untranslated
  • RNA, Transfer
  • Alanine