Directed evolution of DNA or RNA polymerases

The Institut Pasteur is a large scientific organisation located in central Paris with a worldwide reputation in Biology and Microbiology.
It is well connected to public transportation, including Metro lines and trains to south and south-west suburbs.
It is made of 12 Departments and 4 technological or translational Centers that include key technological platforms.
The Department of Structural Biology and Chemistry includes about 150 people: researchers, post-docs, engineers, technicians and students.

Keywords: Directed Evolution, DNA Polymerases, Modified Nucleotides, Microfluidics, Deep Mutational Scanning

DNA polymerases are exquisite catalysts able to synthesize long chains of nucleotides. The current need in biotechnologies for efficient tools to polymerize modified or non-natural nucleic acids motivates numerous enzyme engineering projects all around the world, bringing more and more cutting-edge solutions to this competitive and fast-evolving domain. 

In this project, we propose to use two main state-of-the-art microfluidic-based directed evolution methods [1][2] to engineer polymerases and make them accept modified nucleotides. These ultra-high-throughput selective processes, coupled to NGS technologies, allow to explore efficiently the vast mutational landscape, increasing the chance to find beneficial mutations cycles after cycles.

We are looking for a molecular biologist or biochemist with a keen interest in learning ultra-high-throughput microfluidic droplets generation and sorting, or a microfluidics practitioner keen to apply her/his knowledge to enzyme evolution, or a directed evolution practitioner interested in microfluidic-based high-throughput approaches, all with a PhD degree in one of the aforementioned fields.

The successful candidate will be part of a dynamic team, hosted at the heart of Paris in a very motivating environment and surrounded by several technological platforms bringing a great support to research projects. Beyond the discovery of numerous cutting-edge skills and techniques, the outcomes of the work described above could be both patents and publications shared between the different participants.

[1] Ghadessy et al., PNAS, 2001

[2] Vallejo et al., ACS Syn. Biol., 2019

We are looking for a molecular biologist or biochemist with experience in enzyme evolution and interest in ultra-high-throughput microfluidic droplets generation and sorting, or a microfluidics practitioner keen to apply her/his knowledge to enzyme evolution, or a directed evolution practitioner interested in microfluidic-based high-throughput approaches, all with a PhD degree in one of the aforementioned fields.
 

This CDD is for two years, but an extension by 1 year is possible.