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High-throughput functional genomics: A (myco)bacterial perspective

期刊

MOLECULAR MICROBIOLOGY
卷 -, 期 -, 页码 -

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WILEY
DOI: 10.1111/mmi.15103

关键词

CRISPRi; Mycobacterium tuberculosis; Tn-seq; transposon mutagenesis; TraSH

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Advances in sequencing technologies have provided new insights into bacterial genome composition and dynamics, but the slow validation of inferred genetic function presents a challenge. This is particularly relevant for Mycobacterium tuberculosis, which still has many genes of unknown function despite being one of the first genomes to be sequenced. We summarize the evolution of high-throughput functional genomics and discuss the potential of CRISPR interference in studying bacterial gene function.
Advances in sequencing technologies have enabled unprecedented insights into bacterial genome composition and dynamics. However, the disconnect between the rapid acquisition of genomic data and the (much slower) confirmation of inferred genetic function threatens to widen unless techniques for fast, high-throughput functional validation can be applied at scale. This applies equally to Mycobacterium tuberculosis, the leading infectious cause of death globally and a pathogen whose genome, despite being among the first to be sequenced two decades ago, still contains many genes of unknown function. Here, we summarize the evolution of bacterial high-throughput functional genomics, focusing primarily on transposon (Tn)-based mutagenesis and the construction of arrayed mutant libraries in diverse bacterial systems. We also consider the contributions of CRISPR interference as a transformative technique for probing bacterial gene function at scale. Throughout, we situate our analysis within the context of functional genomics of mycobacteria, focusing specifically on the potential to yield insights into M. tuberculosis pathogenicity and vulnerabilities for new drug and regimen development. Finally, we offer suggestions for future approaches that might be usefully applied in elucidating the complex cellular biology of this major human pathogen.

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