Journal
/
Immunology and Infection
/
Determination of Chemical Inhibitor Efficiency against Intracellular Toxoplasma Gondii Growth Using a Luciferase-Based Growth Assay
JoVE Journal
Immunology and Infection
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Immunology and Infection
Determination of Chemical Inhibitor Efficiency against Intracellular Toxoplasma Gondii Growth Using a Luciferase-Based Growth Assay

Determination of Chemical Inhibitor Efficiency against Intracellular Toxoplasma Gondii Growth Using a Luciferase-Based Growth Assay

DOI:
10.3791/60985-v

•

09:49 min

•

April 29, 2020

•

, , , , ,
1Department of Biological Sciences,Clemson University, 2Eukaryotic Pathogens Innovation Center,Clemson University

Chapters

  • 00:04Introduction
  • 00:49Luciferase-Based ToxoplasmaGrowth Assay
  • 02:25Chemical Compound Inhibition Efficacy Evaluation
  • 04:08CRISPR-Cas9-Based Gene Deletion
  • 05:59Toxoplasma Transfection
  • 06:51Knockout Parasite Cloning
  • 08:06Results: Representative LHVS Inhibition Efficacy Against Intracellular Toxoplasma Growth
  • 09:22Conclusion

Summary

Automatic Translation

Automatic Translation

Presented here is a protocol to evaluate the inhibition efficacy of chemical compounds against in vitro intracellular growth of Toxoplasma gondii using a luciferase-based growth assay. The technique is used to confirm inhibition specificity by genetic deletion of the corresponding target gene. The inhibition of LHVS against TgCPL protease is evaluated as an example.

Tags

Keywords: Toxoplasma GondiiLuciferase-based Growth AssayChemical InhibitorDrug TargetCRISPR-Cas9Intracellular PathogenHigh-throughput ScreeningParasite Doubling TimeHuman Foreskin FibroblastsLuciferase SubstrateMicroplate ReaderNormalized Fold-changeLinear Regression

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/60985?article_type=v

Obtaining Highly Purified Toxoplasma gondii Oocysts by a Discontinuous Cesium Chloride Gradient
check_url/60985?article_type=v

Toxoplasma gondii Cyst Wall Formation in Activated Bone Marrow-derived Macrophages and Bradyzoite Conditions
check_url/60985?article_type=v

Transnuclear Mice with Pre-defined T Cell Receptor Specificities Against Toxoplasma gondii Obtained Via SCNT
check_url/60985?article_type=v

A Genetic Screen to Isolate Toxoplasma gondii Host-cell Egress Mutants
check_url/60985?article_type=v

Growth of Mycobacterium tuberculosis Biofilms
check_url/60985?article_type=v

A Parasite Rescue and Transformation Assay for Antileishmanial Screening Against Intracellular Leishmania donovani Amastigotes in THP1 Human Acute Monocytic Leukemia Cell Line
check_url/60985?article_type=v

Staphylococcus aureus Growth using Human Hemoglobin as an Iron Source
check_url/60985?article_type=v

Genetic Manipulation in Δku80 Strains for Functional Genomic Analysis of Toxoplasma gondii
check_url/60985?article_type=v

Forward Genetics Screens Using Macrophages to Identify Toxoplasma gondii Genes Important for Resistance to IFN-γ-Dependent Cell Autonomous Immunity
check_url/60985?article_type=v

Pairwise Growth Competition Assay for Determining the Replication Fitness of Human Immunodeficiency Viruses

Read Article