Evaluation of Developmental Phenotypes Produced by Morpholino Antisense Targeting of a Sea Urchin Runx Gene

Overview

Journal BMC Biol

Publisher Biomed Central

Specialty Biology

Date 2004 May 11

PMID 15132741

Citations 13

Authors

James A Coffman

Carrie Dickey-Sims

Jeffrey S Haug

John J McCarthy

Anthony J Robertson

Affiliations

Soon will be listed here.

Abstract

Background: Runx transcription factors are important regulators of metazoan development. The sea urchin Runx gene SpRunt was previously identified as a trans-activator of the CyIIIa actin gene, a differentiation marker of larval aboral ectoderm. Here we extend the functional analysis of SpRunt, using morpholino antisense oligonucleotides (morpholinos) to interfere with SpRunt expression in the embryo.

Results: The developmental effects of four different SpRunt-specific morpholinos were evaluated. The two morpholinos most effective at knocking down SpRunt produce an identical mitotic catastrophe phenotype at late cleavage stage that is an artifact of coincidental mis-targeting to histone mRNA, providing a cautionary example of the insufficiency of two different morpholinos as a control for specificity. The other two morpholinos produce gastrula stage proliferation and differentiation defects that are rescued by exogenous SpRunt mRNA. The expression of 22 genes involved in cell proliferation and differentiation was analyzed in the latter embryos by quantitative polymerase chain reaction. Knockdown of SpRunt was found to perturb the expression of differentiation markers in all of the major tissue territories as well as the expression of cell cycle control genes, including cyclin B and cyclin D.

Conclusions: SpRunt is essential for embryonic development, and is required globally to coordinate cell proliferation and differentiation.

Citing Articles

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