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New tools for synthetic biology in Saccharomyces cerevisiae

Grant number: 19/17876-2
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): January 01, 2020
Effective date (End): December 31, 2020
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Elizabeth Bilsland
Grantee:Helena Martins Ribeiro da Cunha Freire
Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Associated research grant:15/03553-6 - Engineering yeast cells for drug discovery, AP.JP


Drugs targeting the central nervous system often fail clinical trials because of their poor transport to the brain. Therefore, our group is interested in unraveling the mechanisms of import of compounds through plasma membrane transporters of the blood-brain barrier. Using genetically modified Saccharomyces cerevisiae as a biological platform, recent studies have shown that it is possible to express human transporters in its plasma membrane. However, the difference in steroid composition between human and yeast plasma membrane limits the correct location of heterologous transporters. Studies indicate that the synthesis of cholesterol in yeast promotes greater success in the expression of human transmembrane proteins. Our present work aims to generate yeasts with adjustable ergosterol synthesis aiming at more efficient incorporation of human proteins in the S. cerevisiae plasma membrane. To this end, we will incorporate a series of synthetic promoters employing CRISPR / Cas9 into the yeast genome. These promoters will initially control the expression of different fluorescent proteins, which can be easily monitored. When optimal circuit regulation is in place, we will develop strains that allow alternation between ergosterol and cholesterol synthesis, allowing it to incorporate yeast and human proteins into its plasma membrane.

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Scientific publications
(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)
ALMEIDA, LUDIMILA DIAS; SILVA, ALI SALIM FARAJ; MOTA, DANIEL CALIXTO; VASCONCELOS, ADRIELLE AYUMI; CAMARGO, ANTONIO PEDRO; PIRES, GABRIEL SILVA; FURLAN, MONIQUE; FREIRE, HELENA MARTINS RIBEIRO DA CUNHA; KLIPPEL, ANGELICA HOLLUNDER; SILVA, SUELEN FERNANDES; et al. Yeast Double Transporter Gene Deletion Library for Identification of Xenobiotic Carriers in Low or High Throughput. MBIO, v. 12, n. 6, . (17/13015-7, 19/14146-3, 17/01986-8, 18/04240-0, 18/05328-8, 18/16672-1, 15/03553-6, 19/17876-2)

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