Screening and characterization of hypoxia responsive promoters using functional genomics

• Grant number: 19/04604-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: April 01, 2019
• End date: February 29, 2020
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Rafael Silva Rocha
• Grantee: Ana Beatriz Faulborn Denardi
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 12/22921-8 - Synthetic biology approaches for deciphering the logic of signal integration in complex bacterial promoters, AP.JP

Abstract

One of the major limitations in the approaches of synthetic biology is the lack of a great diversity of finely characterized biological parts that allow the construction of complex synthetic circuits. On the other hand, the use of bacteria as oncolytic agents is one of the innovative approaches in the treatment of cancer. Some bacterial strains accumulate and grow preferentially within the tumor tissue, indicating a microenvironment favoring the survival and / or growth of some bacterial species and strains. Thus, in the present project, we will mining and characterize new hypoxia responsive promoters from the genome of clinically relevant bacteria Salmonella spp. Thus, a genomic library will be constructed from the genetic material of Salmonella spp. in a vector containing the promoter-free FbFP reporter gene (oxygen-independent fluorescent protein). After the introduction of the library into Escherichia coli, a screening will be performed by promoters that respond to hypoxia conditions. Next, the plasmid DNA of the positive clones will be sequenced in order to identify the novel promoters. Finally, the novel hypoxia responsive promoters will be characterized in E. coli and Salmonella strains. The data generated in this project will be integrated with those obtained in the project that aims at the construction of a genetic circuit to improve the antitumor effect of Salmonella spp.