ZFIN ID: ZDB-PUB-191111-9
Zebrafish etv2 knock-in line labels vascular endothelial and blood progenitor cells
Chestnut, B., Sumanas, S.
Date: 2019
Source: Developmental dynamics : an official publication of the American Association of Anatomists   249(2): 245-261 (Journal)
Registered Authors: Sumanas, Saulius
Keywords: CRISPR, Cas9, RNA-seq, myeloid, red blood cell, transcriptome, vascular endothelial, zebrafish
Microarrays: GEO:GSE139108
MeSH Terms:
  • Animals
  • CRISPR-Associated Protein 9/genetics
  • CRISPR-Associated Protein 9/metabolism
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics
  • Embryo, Nonmammalian/metabolism
  • Gene Expression Regulation, Developmental/genetics
  • Gene Expression Regulation, Developmental/physiology
  • Transcription Factors/genetics
  • Transcription Factors/metabolism
  • Transcriptome/genetics
  • Zebrafish/embryology*
  • Zebrafish/metabolism*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
PubMed: 31705559 Full text @ Dev. Dyn.
ETS transcription factor Etv2 / Etsrp is one of the earliest markers for vascular and hematopoietic progenitors and functions as a key regulator of hematovascular development in multiple vertebrates, including zebrafish. Therefore transgenic etv2 reporter lines provide a valuable tool to study vasculogenesis and hematopoiesis. However, previously generated zebrafish reporter lines do not fully recapitulate the endogenous pattern of etv2 expression.
Here we used CRISPR / Cas9-mediated homology-independent DNA repair approach to knock-in a Gal4 transcriptional activator into the zebrafish etv2 genomic locus, thus generating etv2ci32Gt gene trap line. etv2ci32Gt ; UAS:GFP embryos show GFP expression in vascular endothelial, myeloid and red blood cells. Because gal4 insertion interrupts the etv2 locus, homozygous etv2ci32Gt embryos display defects in vasculogenesis and myelopoiesis, and enable visualizing etv2-deficient hematovascular progenitors in live embryos. Furthermore, we performed differential transcriptome analysis of sorted GFP-positive cells from heterozygous and homozygous etv2ci32Gt embryos. Approximately 500 downregulated genes were identified in etv2ci32Gt homozygous embryos, which include multiple genes expressed in vascular endothelial and myeloid cells.
The etv2ci32Gt gene trap line and the datasets of misregulated genes will be valuable resources to study hematopoietic and vascular development. This article is protected by copyright. All rights reserved.