PgmNr Z615: Positional cloning of cloche, a gene that drives endothelial and hematopoietic lineage specification.

Authors:
Didier Y. R. Stainier ^1,2 ; Sven Reischauer ^1,2 ; Oliver Stone ^1,2 ; Alethia Villasenor ^1,2 ; Suk-Won Jin ¹ ; Neil Chi ¹ ; Marcel Martin ³ ; Miler T. Lee ⁴ ; Michele Marass ² ; Ian Fiddes ¹ ; Taiyi Kuo ¹ ; Won-Suk Chung ¹ ; Sherveen Salek ¹ ; Robert Lerrigo ¹ ; Jessica Alsiö ¹ ; Shujun Luo ⁶ ; Björn Nystedt ⁷ ; Antonio J. Giraldez ⁴ ; Gary P. Schroth ⁵ ; Olov Andersson ⁵

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Institutes
1) Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, California, USA; 2) Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany; 3) Science for Life Laboratory, Dept of Biochemistry and Biophysics, Stockholm University, Solna, Sweden; 4) Department of Genetics, Yale University School of Medicine, New Haven, Connecticut, USA; 5) Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden; 6) Illumina, San Diego, CA, USA; 7) Science for Life Laboratory, Dept of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden.

Abstract:

Vascular and hematopoietic cells organize into specialized tissues during early embryogenesis to supply essential nutrients to all organs and thus play critical roles in development and disease.  At the top of the hemato-vascular specification cascade lies cloche, a gene that when mutated in zebrafish leads to the striking phenotype of loss of most endothelial and hematopoietic cells with a substantial increase in overall cardiomyocyte numbers.  While this mutant has been analyzed extensively to investigate mesoderm diversification and differentiation and continues to be broadly utilized as a unique avascular model, the isolation of the cloche gene has been challenging due to its telomeric location.  Using a deletion allele of cloche, we identified several new cloche candidate genes within this genomic region, and systematically genome-edited each candidate.  Through this exhaustive interrogation, we have finally identified the cloche gene and will report on its identity and further functional analysis.