Development of a photo-activated kill switch device for biocontainment of genetically modified microorganisms

• Grant number: 18/24733-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2019
• End date: April 30, 2021
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Danielle Biscaro Pedrolli
• Grantee: Victor Nunes de Jesus
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Since the emergence of genetic engineering, around 1973, the scientific community has been wondering what the limits of using this technology are. Recently, synthetic biology brought the idea of using cells as biological machines capable of feeling, processing and responding in a programmed way to a given situation. However, the new paradigm breaks with the traditional use of engineered microbes in closed and controlled environments. Synthetic biologists are creating cells as dynamic devices that can interact with the environment or the human body. That rises the need for a high-efficiency and well-characterized biocontainment device in order to avoid spreading of recombinant DNA into the environment. Available devices used in biological reactors, such as auxotrophy, have been presenting excellent results. However, for applications outside the reactor, this solution can be easily bypassed by evolutionary mechanisms (such as mutation and horizontal transfer). Moreover, it does not solve the problem of dispersion of recombinant genetic material. Among the available alternatives, nuclease-based kill switches stand out as highly effective, to the point of being highly toxic to the cells, which could make their application unfeasible. Therefore, this work aims to construct and characterize a genetic circuit using the Cas9 endonuclease associated with photo-activatable negative magnet (nMag) and positive Magnet (pMag) domains as a kill switch alternative for application outside of the bioreactor.

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)

ZOCCA, VITORIA FERNANDA BERTOLAZZI; CORREA, GRACIELY GOMES; LINS, MILCA RACHEL DA COSTA RIBEIRO; DE JESUS, VICTOR NUNES; TAVARES, LEONARDO FERRO; AMORIM, LAURA ARAUJO DA SILVA; KUNDLATSCH, GUILHERME ENGELBERTO; PEDROLLI, DANIELLE BISCARO. The CRISPR toolbox for the gram-positive model bacterium Bacillus subtilis. CRITICAL REVIEWS IN BIOTECHNOLOGY, OCT 2021. (18/24733-0, 14/17564-7, 20/16058-1)