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A Mycobacterial Systems Resource for the Research Community.

J A Judd1, J Canestrari1, R Clark1

  • 1Wadsworth Center, New York State Department of Health, Albany, New York, USA.

Mbio
|March 3, 2021
PubMed
Summary

A new Mycobacterial Systems Resource (MSR) aids functional studies of conserved mycobacterial proteins. This resource, using Mycobacterium smegmatis, provides biological and bioinformatic tools to accelerate research on important mycobacterial species.

Keywords:
CRISPRi clonesMycobacteriumconserved mycobacterial proteinsknockout libraryprotein localization

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Area of Science:

  • Microbiology and Molecular Biology
  • Genomics and Bioinformatics
  • Infectious Diseases

Background:

  • Functional characterization of bacterial proteins, especially in difficult-to-grow species like mycobacteria, lags behind identification.
  • Mycobacterial diseases pose a significant global health burden, yet experimental challenges hinder research into their core functions.
  • Existing model systems like E. coli and B. subtilis are not ideal for studying the conserved biology of diverse mycobacteria.

Purpose of the Study:

  • To establish a comprehensive Mycobacterial Systems Resource (MSR) for facilitating functional characterization of conserved mycobacterial proteins.
  • To provide essential biological and bioinformatic tools applicable to a wide range of mycobacterial species, including pathogens.
  • To accelerate genome annotation and functional assays for mycobacteria by overcoming experimental limitations.

Main Methods:

  • Utilized Mycobacterium smegmatis as a model organism for developing the resource.
  • Created a biological resource including expression plasmid libraries (fluorescent protein fusions), precise gene deletion libraries, and CRISPR-interference (CRISPRi) plasmids.
  • Developed a bioinformatic resource with gene information and protein localization data, focusing on 1,153 highly conserved core genes.

Main Results:

  • Established a resource comprising 1,116 gene-fluorescent protein fusions, 569 precise gene deletions, and 843 CRISPRi plasmids targeting core genes.
  • Generated a bioinformatic database with detailed information on individual genes and protein localization assessments.
  • The resource focuses on 1,153 highly conserved genes common to many mycobacteria, including Mycobacterium tuberculosis.

Conclusions:

  • The Mycobacterial Systems Resource (MSR) provides crucial tools and data to overcome experimental hurdles in mycobacterial research.
  • The MSR is expected to significantly accelerate functional studies of conserved mycobacterial proteins across diverse species.
  • Availability of the MSR (https://msrdb.org/) will benefit the broader mycobacterial research community, aiding studies on pathogens like M. tuberculosis.