Expression of exogenous mRNAs to study gene function in echinoderm embryos.
With the completion of the genome sequencing projects, a new challenge for developmental biologists is to assign a function to the thousands of genes identified. Expression of exogenous mRNAs is a powerful, versatile and rapid technique that can be used to study gene function during development of the sea urchin. This chapter describes how this technique can be used to analyze gene function in echinoderm embryos, how it can be combined with cell transplantation to perform mosaic analysis and how it can be applied to identify downstream targets genes of transcription factors and signaling pathways. We describe specific examples of the use of overexpression of mRNA to analyze gene function, mention the benefits and current limitations of the technique and emphasize the importance of using different controls to assess the specificity of the effects observed. Finally, this chapter details the different steps, vectors and protocols for in vitro production of mRNA and phenotypic analysis.
['Molina|Maria Dolores|MD|', 'Gache|Christian|C|', 'Lepage|Thierry|T|']
D000818:Animals D004625:Embryo, Nonmammalian D018507:Gene Expression Regulation, Developmental D053263:Gene Regulatory Networks D023281:Genomics D012333:RNA, Messenger D012617:Sea Urchins D015398:Signal Transduction D014157:Transcription Factors
mRNA Embryo Sea urchin Mosaic analysis Phenotypic analysis Dominant negative Gain of function In vitro transcription Loss of function Messenger RNA Microinjection