RNAi Screening in Spodoptera frugiperda

Methods Mol Biol. 2016:1470:199-212. doi: 10.1007/978-1-4939-6337-9_16.

Abstract

RNA interference is a potent and precise reverse genetic approach to carryout large-scale functional genomic studies in a given organism. During the past decade, RNAi has also emerged as an important investigative tool to understand the process of viral pathogenesis. Our laboratory has successfully generated transgenic reporter and RNAi sensor line of Spodoptera frugiperda (Sf21) cells and developed a reversal of silencing assay via siRNA or shRNA guided screening to investigate RNAi factors or viral pathogenic factors with extraordinary fidelity. Here we describe empirical approaches and conceptual understanding to execute successful RNAi screening in Spodoptera frugiperda 21-cell line.

Keywords: Insect RNAi; RNA interference; RNAi screening; Sf21 cells; Spodoptera frugiperda; siRNA screening.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Baculoviridae / genetics
  • Cell Line
  • Genes, Reporter
  • Genome, Insect
  • Green Fluorescent Proteins / genetics
  • High-Throughput Screening Assays / instrumentation
  • High-Throughput Screening Assays / methods*
  • RNA Interference*
  • RNA, Small Interfering
  • Reproducibility of Results
  • Spodoptera / genetics*

Substances

  • RNA, Small Interfering
  • Green Fluorescent Proteins