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.
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Acknowledgments
We thank Naoyuki Inagaki for the helpful advice about cell dissociation methods. This work was partly supported by the National BioResource Project (to KK), Grant-in-Aids for Scientific Research on Innovative Areas “Genome Science” (221S0002 to YS), and Grant-in-Aids for Scientific Research (A) (23241063 and 15H02370 to KK) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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Tanabe, H. et al. (2016). Fluorescence-Activated Cell Sorting and Gene Expression Profiling of GFP-Positive Cells from Transgenic Zebrafish Lines. In: Kawakami, K., Patton, E., Orger, M. (eds) Zebrafish. Methods in Molecular Biology, vol 1451. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3771-4_7
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DOI: https://doi.org/10.1007/978-1-4939-3771-4_7
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