Banca de DEFESA: Ana Laura Alfonso Pérez

Uma banca de DEFESA de DOUTORADO foi cadastrada pelo programa.
STUDENT : Ana Laura Alfonso Pérez
DATE: 09/03/2023
TIME: 09:30
LOCAL: Auditório 3
TITLE:

Development of optogenetics-based systems for regulation of gene expression in yeast and the use of serine integrases

KEY WORDS:

yeast, synthetic biology, regulation, optogenetics, integrases

PAGES: 161
BIG AREA: Ciências Biológicas
AREA: Biologia Geral
SUMMARY:

The yeast Komagataella phaffii (Pichia pastoris) has been used as a platform for heterologous protein production for over 20 years because it has high levels of gene expression using the methanol-induced AOX1 (PAOX1) gene promoter, among other advantages. However, methanol, the chemical inducer, is a flammable solution. In recent years, optogenetics has become a widely used tool for the regulation of biological processes such as gene expression, protein localization, signal transduction and proteinprotein interactions. Optogenetics is based on the use of light, a physical inducer, to construct regulable systems. The application of an optogenetic circuit in K. phaffii for the regulation of gene expression allows the use of an induced system that neither affects the yeast metabolism nor suffers interference from cellular metabolism. In the present project, we will seek for the first time to study an optogenetic circuit in yeast in our laboratory. The optogenetic system obtained is still under adjustment, but this first approach brings us closer to the study of optogenetics in our laboratory. Another regulatory system is based on the use of integrases, a class of viral proteins that mediate phage integration and excision into the genome during the lysogenic and lytic life cycle. Recently, 11 orthologous integrases have been characterized and there are no reports of their use in circuit construction in the yeast Saccharomyces cerevisiae to date. This yeast is recognized as a model for studying the biology of eukaryotic organisms, and is one of the most studied and characterized microorganisms. Integrase 13 was evaluated, together with integrase 4 under the control of the galactose responsive promoter. These integrases showed the ability to activate the genetic device. Therefore, these integrases are shown to be functional in yeast, paving the way for the use of integrases as new molecular tools for the development and construction of synthetic circuits. Finally, the regulation system with the integrases was combined with the optogenetic system in order to have a dual regulated system. The final system obtained was functional and adjustments need to be made in the red light/dark conditions.

BANKING MEMBERS:
Externa à Instituição - Milca Rachel da Costa Ribeiro Lins - UFABC
Presidente - 1193623 - FERNANDO ARARIPE GONCALVES TORRES
Interna - 1207115 - ILDINETE SILVA PEREIRA
Externo à Instituição - JOÃO RICARDO MOREIRA DE ALMEIDA - EMBRAPA
Interno - 2122537 - MARCELO DE MACEDO BRIGIDO