en
Book chapter
Open access
English

Combining RNAi-Mediated β-Catenin Inhibition and Reaggregation to Study Hydra Whole-Body Regeneration

PublisherNew-York : Humana Press
Collection
  • Methods in molecular biology; 2450
Publication date2022-03-31
First online date2022-04-01
Abstract

In addition to its ability to regenerate any amputated body part, the Hydra freshwater polyp shows the amazing ability to regenerate as a full polyp after a complete dissociation of its tissues. The developmental processes at work in reaggregates undergoing whole-body regeneration can be investigated at the molecular level by RNA interference (RNAi). Here we provide a protocol that combines β-catenin RNAi with reaggregation. This protocol serves as a basis to generate "RNAi-reaggregates," followed by the extraction of high-quality RNA for the precise quantification of gene expression by real-time PCR. This protocol is efficient, providing both a molecular signature, with the significant downregulation of β-catenin and Wnt3, as well as a robust phenotype, the lack of axis formation, which is observed in all reaggregates.

eng
Keywords
  • Gene knockdown
  • Hydra
  • Patterning
  • Reaggregation
  • Whole-body regeneration
  • Wnt3
  • QPCR
  • SiRNA electroporation
  • Β-catenin
Research group
Citation (ISO format)
VOGG, Matthias Christian, GALLIOT, Brigitte. Combining RNAi-Mediated β-Catenin Inhibition and Reaggregation to Study Hydra Whole-Body Regeneration. In: Whole-Body Regeneration: Methods and Protocols. New-York : Humana Press, 2022. p. 635–647. (Methods in molecular biology) doi: 10.1007/978-1-0716-2172-1_34
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Book chapter (Published version)
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Technical informations

Creation04/02/2022 11:55:00 AM
First validation04/02/2022 11:55:00 AM
Update time03/16/2023 6:20:26 AM
Status update03/16/2023 6:20:26 AM
Last indexation05/06/2024 10:35:29 AM
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