NIH U19 FLUTE Project
2013-2018
Project Goals
Vast quantities of new genome sequence are added to public databases
on a daily basis. But, while we are constantly deluged with new gene
sequences we still have a limited ability to define their
functions. Almost all functions are defined by comparison with genes
from other strains in which experimental data are available. But these
experimental data have not kept pace with the availability of new
sequence. In fact, for most bacterial species, many genes have no
known function and even those that are annotated have only limited
information. This is particularly striking for Mycobacterium
tuberculosis (Mtb), where only 52% of protein-coding genes have a
putative function.
We plan to discover the roles of genes from Mtb, an important and
widespread human pathogen, with previously unknown
functions. Critically, we will target genes that fulfill vital roles
in bacterial growth and survival, genes we have previously identified
in genome-wide screens. This offers three major advantages. First, we
will concentrate on only the most important unannotated genes. Second,
phenotypes allow us to use the power of synthetic lethality to
identify interactions. And third, phenotypes provide us with a context
in which we can understand the outcome of biochemical and genetic
assays. In addition to defining the roles for Mtb genes we aim to
establish an efficient pathway for identifying gene function that can
serve as a paradigm for other bacterial species. To accomplish this we
will undertake an ambitious program to construct large numbers of Mtb
mutants. This will be possible as we will take advantage of
substantial mycobacterial genetic expertise among the
participants. Moreover, we will use a number of analytic modalities
brought in through a set of highly interconnected projects and cores.
Funding
This project is funded by the NIH as part of the NIAID Functional
Genomics Program, under grant U19 AI107774. The NIAID Functional
Genomics Program for understanding the functions of uncharacterized
genes in infectious disease pathogens will generate experimental data
to determine the biochemical function(s) of hypothetical genes,
unknown open reading frames, and noncoding RNAs. The program will
apply state-of-the-art technologies to determine the biochemical and
physiological roles of these gene components. Obtaining a more
comprehensive understanding of uncharacterized genes in infectious
disease pathogens will lead to improved genomic annotation and allow
for the development of potential new targets for medical diagnostics,
therapeutics and vaccines. The program will distribute data, software,
and reagents generated from the research projects to the broader
scientific community.
The successor of the original project has been funded by NIH as P01 AI143575 (Pathway Analysis in Tuberculosis; program director: S. Ehrt; 2020-2025).
Investigators
Eric Rubin, Harvard School of Public Health
Sarah Fortune, Harvard School of Public Health
Dirk Schnappinger, Weill Cornell Medical College, NY
Sabine Ehrt, Weill Cornell Medical College, NY
Kyu Y Rhee, Weill Cornell Medical College, NY
Chris Sassetti, University of Massachusetts Medical School
Thomas Ioerger, Texas A&M University
James Sacchettini, Texas A&M University
Steven A Carr, Broad Institute
Jeremy Rock, Rockefeller University
Scarlet Shell, Worcester Polytechnic Institute
Targets
Techniques
Software
- TRANSIT - python-based software for analysis TnSeq data (from Tom Ioerger at Texas A&M)
- RockHopper - java-based software for analyzing RNAseq data (from Brian Tjaden at Wellesley College)