Flow Cytometry-Based Cell Type-Specific Assessment of Target Regulation by Pulmonary siRNA Delivery

Methods Mol Biol. 2019:1943:365-375. doi: 10.1007/978-1-4939-9092-4_24.

Abstract

Pulmonary siRNA delivery has attracted strong interest and has been reported to successfully mediate target gene knockdown in a number of disease models. However, the nature of the epithelial cells that eventually take up siRNA and the question if other lung cell types may also be transfected has so far been neglected. Therefore, we describe here a flow cytometry-based method using transgenic enhanced green fluorescence protein (EGFP) expressing mice for the differentiation of transfected lung cell populations based on their antigen expression.

Keywords: Differentiation; EGFP mice; Epithelial cells; Flow cytometry; Knockdown; Lung; Type II pneumocytes; siRNA delivery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bronchoalveolar Lavage / methods
  • Cell Separation / methods
  • Female
  • Flow Cytometry / methods
  • Gene Knockdown Techniques / methods*
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • Lung / cytology
  • Lung / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Nanoparticles / chemistry
  • RNA Interference
  • RNA, Small Interfering / genetics*
  • Transfection / methods*

Substances

  • RNA, Small Interfering
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins