Publications by authors named "Mei-Yi Yan"

Tuberculosis (TB), the leading cause of death from bacterial infections worldwide, results from infection with Mycobacterium tuberculosis (Mtb). The antitubercular agents delamanid (DLM) and pretomanid (PMD) are nitroimidazole prodrugs that require activation by an enzyme intrinsic to Mtb; however, the mechanism(s) of action and the associated metabolic pathways are largely unclear. Profiling of the chemical-genetic interactions of PMD and DLM in Mtb using combined CRISPR screening reveals that the mutation of rv2073c increases susceptibility of Mtb to these nitroimidazole drugs both in vitro and in infected mice, whereas mutation of rv0078 increases drug resistance.

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CRISPR screening, including CRISPR interference (CRISPRi) and CRISPR-knockout (CRISPR-KO) screening, has become a powerful technology in the genetic screening of eukaryotes. In contrast with eukaryotes, CRISPR-KO screening has not yet been applied to functional genomics studies in bacteria. Here, we constructed genome-scale CRISPR-KO and also CRISPRi libraries in (Mtb).

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Toxin-antitoxin (TA) systems are ubiquitous genetic elements in prokaryotes, but their biological importance is poorly understood. Mycobacterium smegmatis contains eight putative TA systems. Previously, seven TAs have been studied, with five of them being verified as functional.

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Multidrug-resistant () infection seriously endangers global human health, creating an urgent need for new treatment strategies. Efficient genome editing tools can facilitate identification of key genes and pathways involved in bacterial physiology, pathogenesis, and drug resistance mechanisms, and thus contribute to the development of novel treatments for drug-resistant . Here, we report a two-plasmid system, , used to inactivate genes and introduce point mutations in .

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New tools for genetic manipulation of are needed for the development of new drug regimens and vaccines aimed at curing tuberculosis infections. Clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated protein (Cas) systems generate a highly specific double-strand break at the target site that can be repaired via nonhomologous end joining (NHEJ), resulting in the desired genome alteration. In this study, we first improved the NHEJ repair pathway and developed a CRISPR-Cas-mediated genome-editing method that allowed us to generate markerless deletion in , , and Then, we demonstrated that this system could efficiently achieve simultaneous generation of double mutations and large-scale genetic mutations in Finally, we showed that the strategy we developed can also be used to facilitate genome editing in The global health impact of necessitates the development of new genetic tools for its manipulation, to facilitate the identification and characterization of novel drug targets and vaccine candidates.

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Article Synopsis
  • CRISPR-Cas12a (Cpf1) is a novel and effective tool for genome editing that enhances genetic manipulation in bacteria by allowing precise modifications like point mutations, deletions, and gene replacements.
  • * The development of a CRISPR-Cas12a-assisted recombineering system enables efficient creation of markerless mutations without the need for antibiotic resistance genes, making it a cleaner alternative to traditional methods.
  • * This system's capabilities include targeting different sites compared to CRISPR-Cas9, which improves precision in editing and can also be applied to manipulate plasmids, providing a comprehensive approach for genetic research and applications in bacteria.
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