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Forging technical breakthroughs for colonial ascidian developmental biology research: transgenesis and mutagenesis in Botryllus schlosseri

Grant number: 19/06927-5
Support Opportunities:Scholarships abroad - Research
Start date: September 01, 2019
End date: August 31, 2020
Field of knowledge:Biological Sciences - Morphology - Embryology
Principal Investigator:Federico David Brown Almeida
Grantee:Federico David Brown Almeida
Host Investigator: Stefano Tiozzo
Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Institution abroad: Laboratoire de Biologie du Développement de Villefranche-sur-Mer (LBDV), France  
Associated research grant:15/50164-5 - Stem cells, regeneration, and the evolution of coloniality in ascidians, AP.JP

Abstract

For nearly sixty years, colonial ascidians (mainly Botryllus schlosseri) have been used as laboratory models to study chordate origins, including the evolution of the immune system and allorecognition, as well as different aspects of colonial animal biology, including asexual reproduction, budding (i.e. blastogenesis), regeneration, or adult somatic and germline stem cell transmission and parasitism. Despite the vast amount of developmental biology research in B. schlosseri, protocols for transgenesis and mutagenesis have fallen short hampering studies of gene regulation and function. Colonial ascidians have internal fertilization and brood their embryos impeding easy access to the eggs or 1-cell embryo for electroporation and microinjection. For this proposal, I plan to generate a protocol for transgenesis in colonial ascidians by: (i) microinjection of gene construct directly into colony eggs or 1-cell brooding embryos, and (ii) electroporation of circulatory germline stem cells in colony extracts; and I plan to generate knockout colonial ascidian mutant lines by: (iii) a self-fertilization mutant screen, (iv) an ENU or radiation based forward genetic screen, or (iv) CRISPR for gene targeted knockouts. Future research and new discoveries in developmental biology using colonial ascidians will directly benefit from technical breakthroughs outlined in this proposal. To our knowledge, this is the first attempt to generate stable transgenic and mutant lines for any colonial animal, and the Tiozzo laboratory is currently at the forefront of developing these tools. (AU)

News published in Agência FAPESP Newsletter about the scholarship:
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Scientific publications (5)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MOREANO-ARROBO, LOLA; PEREZ, OSCAR D.; BROWN, FEDERICO D.; OYARZUN, FERNANDA X.; CANALES-AGUIRRE, CRISTIAN B.. The ``Mexican dancer{''} in Ecuador: molecular confirmation, embryology and planktotrophy in the sea slug Elysia diomedea. INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY, v. 65, n. 4-6, 2, p. 323-332, . (18/50017-0, 15/50164-5, 19/06927-5)
HIEBERT, LAUREL S.; VIEIRA, LEANDRO M.; TIOZZO, STEFANO; SIMPSON, CARL; GROSBERG, RICHARD K.; MIGOTTO, ALVARO E.; MORANDINI, ANDRE C.; BROWN, FEDERICO D.. From the individual to the colony: Marine invertebrates as models to understand levels of biological organization. JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION, v. 336, n. 3, . (18/05923-3, 15/50164-5, 19/06927-5, 18/50017-0, 15/14052-8)
BROWN, FEDERICO D.. Evolution of animal coloniality and modularity: Emerging themes. JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION, v. 336, n. 3, p. 4-pg., . (19/06927-5, 18/50017-0, 15/50164-5)
JIMENEZ-MERINO, JUAN; DE ABREU, ISADORA SANTOS; HIEBERT, LAUREL S.; ALLODI, SILVANA; TIOZZO, STEFANO; DE BARROS, CINTIA M.; BROWN, FEDERICO D.. Putative stem cells in the hemolymph and in the intestinal submucosa of the solitary ascidian Styela plicata. EVODEVO, v. 10, n. 1, . (19/06927-5, 16/07607-6, 15/50164-5, 15/14052-8, 18/05923-3)
MOREANO-ARROBO, LOLA; PEREZ, OSCAR D.; BROWN, FEDERICO D.; OYARZUN, FERNANDA X.; CANALES-AGUIRRE, CRISTIAN B.. The "Mexican dancer" in Ecuador: molecular confirmation, embryology and planktotrophy in the sea slug Elysia diomedea. INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY, v. 65, n. 4-6, p. 10-pg., . (19/06927-5, 18/50017-0, 15/50164-5)