Next Generation Sequencing Quantitative Analysis of Wild Type and Shoc2 -/- sox10-positive cells in zebrafish

To determine how Shoc2-mediated signals affect expression of genes controlling specification of neural crest cells in zebrafish development, we applied RNA-sequencing. shoc2 CRISPR/Cas9 mutants that are transgenic for sox10:dsRed – marking neural crest progenitors were utilized in these experiments. mRNAs was isolated from the FACS-purified sox10:dsRed positive cells of shoc2 CRISPR/Cas9 mutant and wild type larvae. This approach allows for a robust and targeted quantitative assessment of how Shoc2-ERK1/2 signaling affects emergence of several neural crest lineages. The differences in the transcriptome of cells purified from shoc2 crispants and control embryos were determined. Overall design: examination of the transcriptional changes in zebrafish larvea that lost shoc2 expression