Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Characterizing Modulators of Protease-Activated Receptors with a Calcium Mobilization Assay Using a Plate Reader07:13

Characterizing Modulators of Protease-Activated Receptors with a Calcium Mobilization Assay Using a Plate Reader

942
An improved protocol for a calcium mobilization assay with endothelial cells, used to identify ligands of protease-activated receptors (PARs), has been developed. The new protocol reduces total assay time by 90-120 min and yields reproducible concentration-response...
942
Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira14:49

Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira

5.7K
Here, the application of CRISPR interference (CRISPRi) for specific gene silencing in Leptospira species is described. Leptospira cells are transformed by conjugation with plasmids expressing dCas9 (catalytically "dead" Cas9) and a single-guide RNA (sgRNA), responsible for base pairing to the desired genomic target. Methods to validate gene silencing are presented.
5.7K
Asthma: Pathogenesis and Management01:20

Asthma: Pathogenesis and Management

1.2K
Asthma is a chronic pulmonary condition involving inflammation of the airways, hyper-reactivity, and reversible obstruction of the airways. This condition can significantly impact a person's quality of life, making breathing difficult and leading to distressing symptoms.
Asthma is classified as allergic and non-allergic. Allergens such as dust mites, pollen, and pet dander trigger allergic asthma, while factors like cold air, intense emotions, or exercise can induce non-allergic asthma.
1.2K
Cystic Fibrosis: Pathogenesis01:23

Cystic Fibrosis: Pathogenesis

735
Cystic fibrosis (CF), an autosomal recessive disorder, significantly affects the function of exocrine glands. This genetically inherited disease is characterized by the production of thick and sticky mucus, which can severely affect various organs and systems in the body.
CF is primarily caused by a genetic mutation in a chromosome 7 gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR) protein. The most common gene mutation leading to CF is the ΔF508 mutation,...
735
Pulmonary Hypertension: Classification and Pathogenesis01:30

Pulmonary Hypertension: Classification and Pathogenesis

586
Pulmonary hypertension (PH) is a severe health condition in which the mean pulmonary arterial pressure increases to 25 mmHg or more, even when the body is at rest. This high pressure in the blood vessels that transport blood from the heart to the lungs can cause various symptoms, including shortness of breath, can lead to right heart failure, and significantly affect the overall quality of life.
There are various classifications for PH, each relating to different underlying causes and also...
586
Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments08:40

Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments

4.8K
Ion mobility spectrometry (IMS) is an interesting complement to mass spectrometry for the characterization of biomolecules, notably because it is sensitive to isomerism. This protocol describes a tandem IMS (IMS/IMS) experiment, which allows the isolation of a molecule and the generation of the mobility profiles of its fragments.
4.8K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Intracellular barriers and receptor masking limit success of a <i>Pseudomonas aeruginosa</i> clinical phage family.

bioRxiv : the preprint server for biology·2026
Same author

Explainable machine learning incorporating CHI3L1 enhances liver fibrosis staging in chronic hepatitis B: The CHILI multicenter study.

Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver·2026
Same author

From microbes to molecules: Gut microbiota as a prerequisite threshold determinant for cancer immunotherapy efficacy.

Microbiological research·2026
Same author

Advancing Antimicrobial Stewardship With Diagnostic Excellence: The Role of Bayesian Reasoning.

Open forum infectious diseases·2026
Same author

Systematic evaluation of TCGA tumor microbiota reveals context-dependent reliability.

mSystems·2026
Same author

Programmable kinetic barcoding for multiplexed RNA detection with Cas13a.

Nature biomedical engineering·2026

Related Experiment Video

Updated: Jan 20, 2026

Author Spotlight: Developing Parmodulins to Target Protease-Activated Receptors for Inflammation Control
07:13

Author Spotlight: Developing Parmodulins to Target Protease-Activated Receptors for Inflammation Control

Published on: May 24, 2024

942

Modulating Pathogenesis with Mobile-CRISPRi.

Jiuxin Qu1,2, Neha K Prasad1,2, Michelle A Yu2

  • 1Chan Zuckerberg Biohub, San Francisco, California, USA.

Journal of Bacteriology
|September 5, 2019
PubMed
Summary
This summary is machine-generated.

Mobile-CRISPRi technology effectively controls conditionally essential (CE) genes in pathogenic bacteria during infection. This tool aids in understanding bacterial virulence and developing new antimicrobial strategies against antibiotic resistance.

Keywords:
CRISPRiPseudomonas aeruginosaconditionally essential genesinfection modeltype III secretionvirulence lifestyle genes

More Related Videos

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira
14:49

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira

Published on: August 14, 2021

5.7K
Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments
08:40

Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments

Published on: January 20, 2022

4.8K

Related Experiment Videos

Last Updated: Jan 20, 2026

Author Spotlight: Developing Parmodulins to Target Protease-Activated Receptors for Inflammation Control
07:13

Author Spotlight: Developing Parmodulins to Target Protease-Activated Receptors for Inflammation Control

Published on: May 24, 2024

942
Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira
14:49

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira

Published on: August 14, 2021

5.7K
Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments
08:40

Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments

Published on: January 20, 2022

4.8K

Area of Science:

  • Microbiology
  • Genetics
  • Infectious Diseases

Background:

  • Conditionally essential (CE) genes are critical for bacterial pathogenesis but challenging to study due to their essential nature during infection.
  • Developing novel antimicrobials requires understanding the precise roles of CE genes in bacterial survival within host environments.
  • Existing genetic tools lack the precision needed to manipulate CE gene expression in vivo, hindering research into bacterial virulence factors.

Purpose of the Study:

  • To demonstrate the efficacy of Mobile-CRISPRi for controlling CE gene expression in a live animal infection model.
  • To optimize Mobile-CRISPRi for use in a murine pneumonia model caused by *Pseudomonas aeruginosa*.
  • To validate Mobile-CRISPRi as a tool for probing CE gene function and identifying new antimicrobial targets.

Main Methods:

  • Development and optimization of a Mobile-CRISPRi system for inducible gene knockdown in *Pseudomonas aeruginosa*.
  • Application of Mobile-CRISPRi in a murine model of lung infection to assess the impact on CE gene expression.
  • Genetic manipulation of the type III secretion system (T3SS) activator ExsA using Mobile-CRISPRi to evaluate its role in virulence.

Main Results:

  • Optimized Mobile-CRISPRi demonstrated effective and tunable knockdown of CE genes in *P. aeruginosa* within a murine pneumonia model.
  • Knockdown of the ExsA gene using Mobile-CRISPRi successfully blocked T3SS effector protein secretion in vitro.
  • Partial gene perturbation of ExsA significantly attenuated bacterial virulence in the mouse infection model.

Conclusions:

  • Mobile-CRISPRi is a powerful and versatile tool for studying conditionally essential virulence genes in bacterial pathogenesis models.
  • This technology enables precise manipulation of gene expression in vivo, facilitating the discovery of novel antimicrobial targets.
  • Mobile-CRISPRi holds significant promise for advancing research in infectious diseases and combating antibiotic resistance.