Expression of exogenous mRNAs to study gene function in echinoderm embryos.
Methods Cell Biol
; 151: 239-282, 2019.
Article
em En
| MEDLINE
| ID: mdl-30948011
ABSTRACT
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Ouriços-do-Mar
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RNA Mensageiro
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Regulação da Expressão Gênica no Desenvolvimento
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Genômica
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Methods Cell Biol
Ano de publicação:
2019
Tipo de documento:
Article
País de afiliação:
França