ZFIN ID: ZDB-PUB-160729-23
Fluorescence-Activated Cell Sorting and Gene Expression Profiling of GFP-Positive Cells from Transgenic Zebrafish Lines
Tanabe, H., Seki, M., Itoh, M., Deepak, A., Lal, P., Horiuchi, T., Suzuki, Y., Kawakami, K.
Date: 2016
Source: Methods in molecular biology (Clifton, N.J.) 1451: 93-106 (Chapter)
Registered Authors: Kawakami, Koichi
Keywords: Enhancer trap, FACS, GFP, Gal4-UAS system, Gene expression analysis, Gene trap, Next-generation sequence, RNA extract, RNA-seq, Tol2 transgenesis
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*
PubMed: 27464803 Full text @ Meth. Mol. Biol.
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.