CRISPR Screening and Comparative LC-MS Analysis Identify Genes Mediating Efficacy of Delamanid and Pretomanid against Mycobacterium tuberculosis.

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.

Corrigendum: Comparison of prognosis between microscopically positive and negative surgical margins for primary gastrointestinal stromal tumors: A systematic review and meta-analysis.

[This corrects the article DOI: 10.3389/fonc.2022.

Mining of the Novel Virulent ATP-Binding Cassette Importers in by Comparative Genomic Strategy.

The virulent ATP-binding cassette (ABC) importers from , the most common native multidrug resistant and emerging opportunistic pathogen in rapidly growing NTM, were explored by comparative genomic study, in view of the fact that the ABC importers of , responsible for uptaking metals, anions, amino acids, peptides, sugars, and other crucial substances from the host, had been proved to be closely related with the bacillus's virulence, survival in the host macrophages, antibiotic resistance, modulation of host immune system, and so on, although detailed mechanism was unclear. For virulent ABC importers from predicted by orthology and phylogeny analysis of nucleotide-binding domains (NBDs) of , , and , the antibiotic susceptibility of overexpression transformant and knockout mutant was assayed after confirmation by experiment. Three-domain importers were dominant ones in (60.

Application of combined CRISPR screening for genetic and chemical-genetic interaction profiling in .

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).

Comparison of Prognosis Between Microscopically Positive and Negative Surgical Margins for Primary Gastrointestinal Stromal Tumors: A Systematic Review and Meta-Analysis.

Background: This meta-analysis aimed to determine the prognostic impact of microscopically positive margins (R1) on primary gastrointestinal stromal tumors.

Methods: A literature search was performed using PubMed, Embase, Web of Science, and Cochrane Library for studies up to 23 November 2020. The pooled disease-free survival (DFS) and overall survival (OS) between R1 and negative margins (R0) were estimated using a random-effects model.

Programmable Base Editing in Using an Engineered CRISPR RNA-Guided Cytidine Deaminase.

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 .

A CRISPR-Assisted Nonhomologous End-Joining Strategy for Efficient Genome Editing in Mycobacterium tuberculosis.

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.