ArticleNucleic acids research2016
Investigating essential gene function in Mycobacterium tuberculosis using an efficient CRISPR interference system.
Article in Nucleic acids research, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 84 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
84 citing papers in PubMed, 165 citations in OpenAlex.
- Mycobacterium intracellulare M.i.198-A Promising Strain for Studying Pathogenesis of Nontuberculous Mycobacterial Pulmonary Disease (NTM-PD).Microbiology and immunology · 2026Review
- Transcriptional control of asparagine utilization regulates mycobacterial pH-driven adaptation at the host-pathogen crossroads.The Journal of biological chemistry · 2026Article
- ClpC1-dependent iron homeostasis underlies mycobacterial defense against oxygen-driven Fenton reaction during reactivation.Microbiology spectrum · 2026Article
- Genetic interference of distinctive Mycobacterium tuberculosis peptidoglycan modifications enhances β-lactam susceptibility and reveals expression-sensitive host immune dynamics.Scientific reports · 2026Article
- Nucleoid-associated protein HU constrains transcription elongation to influence cellular functions, virulence, and drug tolerance in Mycobacterium tuberculosis.Nucleic acids research · 2026Article
- Physiological, genetical and morphological alterations in Mycobacterium avium subsp. paratuberculosis mutants generated with the CRISPRi system.BMC microbiology · 2026Article
- Functional genomics reveals strain-specific genetic requirements conferring hypoxic growth ineLife · 2025Article
- Effects of CwlM, a peptidoglycan synthesis regulator, on beta-lactam tolerance and host-pathogen interactions.BMC microbiology · 2025Article
- Uncovering Insights Into the Biology of Mycobacterium tuberculosis Using Genetic Tools.MicrobiologyOpen · 2025Review
- Functional analysis of the intracellular survival ofJournal of bacteriology · 2025Article
- CRISPRi knockdown of mycobacterial tkt gene potentiates the anti-mycobacterial activity of phyto-compounds from selected medicinal plants.BMC complementary medicine and therapies · 2025Article
- A hybrid machine learning model with attention mechanism and multidimensional multivariate feature coding for essential gene prediction.BMC biology · 2025Article
- The geneAntimicrobial agents and chemotherapy · 2025Article
- Fluorescence-based CRISPR interference system for controlled genetic repression and live single-cell imaging in mycobacteria.FEBS letters · 2025Article
- Growth in biofilms preparesFrontiers in microbiology · 2025Article
- Efficient genome engineering in Mycolicibacterium neoaurum using Cas9 from Streptococcus thermophilus.Biotechnology letters · 2024Article
- The CRISPR-dCas9 interference system suppresses inhA gene expression in Mycobacterium smegmatis.Scientific reports · 2024Article
- EfpA is required for regrowth ofAntimicrobial agents and chemotherapy · 2024Article
- Mycobacterium tuberculosis produces D-serine under hypoxia to limit CD8Nature microbiology · 2024Article
- Article
24 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
Despite many methodological advances that have facilitated investigation of Mycobacterium tuberculosis pathogenesis, analysis of essential gene function in this slow-growing pathogen remains difficult. Here, we describe an optimized CRISPR-based method to inhibit expression of essential genes based on the inducible expression of an enzymatically inactive Cas9 protein together with gene-specific guide RNAs (CRISPR interference). Using this system to target several essential genes of M. tuberculosis, we achieved marked inhibition of gene expression resulting in growth inhibition, changes in susceptibility to small molecule inhibitors and disruption of normal cell morphology. Analysis of expression of genes containing sequences similar to those targeted by individual guide RNAs did not reveal significant off-target effects. Advantages of this approach include the ability to compare inhibited gene expression to native levels of expression, lack of the need to alter the M. tuberculosis chromosome, the potential to titrate the extent of transcription inhibition, and the ability to avoid off-target effects. Based on the consistent inhibition of transcription and the simple cloning strategy described in this work, CRISPR interference provides an efficient approach to investigate essential gene function that may be particularly useful in characterizing genes of unknown function and potential targets for novel small molecule inhibitors.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.