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Cloning and characterization of 10 human blood-brain-barrier transporters

Grant number: 18/05328-8
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2018
Effective date (End): May 31, 2019
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Principal Investigator:Elizabeth Bilsland
Grantee:Gabriel Silva Pires
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

Abstract

Drugs directed to the central nervous system frequently fail in clinical trials because of their poor transport into the brain. This is due to the inefficient import of drugs though the blood brain barrier.In this project, we aim to use the yeast Saccharomyces cerevisiae as a platform for studying the transport of chemicals into and out of cells. To that end the scientific initiation student will contribute to the creation of a collection of plasmids encoding fluorescently tagged human plasma membrane transporters. These will be transformed into yeast strains where the native ergosterol synthesis pathway is replaced for a synthetic human cholesterol synthesis pathway. We have promising preliminary results indicating that strains with cholesterol in their plasma membranes have the ability to efficiently incorporate human trans-membrane proteins. Therefore, strains with a humanized plasma membrane can be hosts for the heterologous expression of blood-brain barrier transporters. Once yeast strains harbouring the human transporters have been constructed, the student will evaluate the subcellular localization of each transporter and perform functional assays to confirm the substrate specificity for each transporter.

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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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