Analysis for the regulatory mechanisms underlying the cranial vascular formation

2016 Fiscal Year Final Research Report

• Project/Area Number: 26460255
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General anatomy (including histology/embryology)
• Research Institution: Iwate Medical University
• Principal Investigator: Kimura Eiji 岩手医科大学, 医学部, 講師 (50405750)
• Co-Investigator(Renkei-kenkyūsha): KAWAHARA Atsuo 山梨大学, 総合研究部, 教授 (10362518)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: ゼブラフィッシュ / CRISPR/Cas9 / 脈管形成 / 血管新生 / 遺伝子破壊

Outline of Final Research Achievements

Previously, we performed microarray analysis of etv2 deficient zebrafish embryos, which was one of the earliest markers for angioblasts. We first identified 165 genes that were up-regulated more than 1.5 fold by silencing etv2 expression, then analyzed their expression patterns at 12-somite stage to evaluate whether these genes were associated with the cranial vascular formation, and finally selected 11 genes for the loss of function assay. In this study, we knocked out these 11 genes using the CRISPR/ Cas9 system to analyze the mechanisms of cerebral vasculature formation. As a result, we succeeded in producing sgRNAs for each gene which was capable of the genome editing. For 10 genes, we established heterozygous zebrafish lines in which targeted genes were knocked out by the frame-shift and analysis of the phenotype induced by loss of function were in progress. Additionally, we succeeded in revealing the primary ocular vasculature formation both in zebrafish and mouse embryos.

Free Research Field

発生学