Overview

Project title
Functional anti-tubercular screening of M. tuberculosis and M. smegmatis by CRISPRi-seq

People involved
Principal investigator : Prof Valerie Mizrahi
Co-investigator: Mandy Mason
Collaborator: Dr Luiz Carvalho (Francis Crick Institute)
Collaborator: Dr Jeremy Rock (Rockefeller University)
Bioinformatics support team: Katie Lennard

Short description of project
'In this project, I aim to identify bacillary factors that mitigate cidality of anti-tubercular drugs by revealing Mtb mutants displaying hypersusceptibility to routinely used and experimental antitubercular drugs with differing modes of action. We aim to utilise high-throughput CRISPRi-seq to perform a chemical-genetic screen to estimate mutant fitness in a comprehensive pooled library of Mtb mutants representing both essential and non-essential genes. This will expose mutants with decreased fitness in response to antitubercular drug treatment over time.'

Objectives
Advice regarding data handling and storage, as well as data processing and analysis:
Mandy has been using web based software (MAGeCk VISPR) but would appreciate advice in this regard.
Mandy currently has data sets for M. smegmatis and is expecting sequencing from experiments in M. tuberculosis in mid-February. She would require assistance to process these.