Fluorescence-Activated Cell Sorting and Gene Expression Profiling of GFP-Positive Cells from Transgenic Zebrafish Lines

Methods Mol Biol. 2016:1451:93-106. doi: 10.1007/978-1-4939-3771-4_7.

Abstract

Gene expression profiling is a useful approach for deeper understanding of the specificity of cells, tissues, and organs in the transcriptional level. Recent development of high-throughput next-generation sequence (NGS) allows the RNA-seq method for this profiling. This method provides precise information of transcripts about the quantitation and the structure such as the splicing variants. In this chapter, we describe a method for gene expression profiling of GFP-positive cells from transgenic zebrafish by RNA-seq. We labeled specific cells in the brain with GFP by crossing a Gal4 driver line with the UAS:GFP line, isolated those cells by fluorescence-activated cell sorting (FACS), and analyzed by RNA-seq.

Keywords: Enhancer trap; FACS; GFP; Gal4-UAS system; Gene expression analysis; Gene trap; Next-generation sequence; RNA extract; RNA-seq; Tol2 transgenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics
  • Animals, Genetically Modified / metabolism*
  • Enhancer Elements, Genetic / genetics*
  • Flow Cytometry / methods*
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Developmental / genetics
  • Gene Expression Regulation, Developmental / physiology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • High-Throughput Nucleotide Sequencing
  • Zebrafish / genetics
  • Zebrafish / metabolism*

Substances

  • Green Fluorescent Proteins