Practical Guide for Ascidian Microinjection: Phallusia mammillata.
Adv Exp Med Biol
; 1029: 15-24, 2018.
Article
em En
| MEDLINE
| ID: mdl-29542077
ABSTRACT
Phallusia mammillata has recently emerged as a new ascidian model. Its unique characteristics, including the optical transparency of eggs and embryos and efficient translation of exogenously introduced mRNA in eggs, make the Phallusia system suitable for fluorescent protein (FP)-based imaging approaches. In addition, genomic and transcriptomic resources are readily available for this ascidian species, facilitating functional gene studies. Microinjection is probably the most versatile technique for introducing exogenous molecules such as plasmids, mRNAs, and proteins into ascidian eggs/embryos. However, it is not practiced widely within the community; presumably, because the system is rather laborious to set up and it requires practice. Here, we describe in as much detail as possible two microinjection methods that we use daily in the laboratory one based on an inverted microscope and the other on a stereomicroscope. Along the stepwise description of system setup and injection procedure, we provide practical tips in the hope that this chapter might be a useful guide for introducing or improving a microinjection setup.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Urocordados
/
RNA Mensageiro
/
Animais Geneticamente Modificados
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Técnicas de Transferência de Genes
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Microinjeções
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Adv Exp Med Biol
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
França