Accedi

Broad Institute of Harvard and MIT

2 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Measuring Growth and Gene Expression Dynamics of Tumor-Targeted S. Typhimurium Bacteria

Tal Danino^*1, Arthur Prindle^*2, Jeff Hasty^2,3,4, Sangeeta Bhatia^1,5,6,7,8

¹Health Sciences and Technology, Massachusetts Institute of Technology, ²Department of Bioengineering, University of California, San Diego , ³Biocircuits Institute, University of California, San Diego , ⁴Molecular Biology Section, Division of Biological Science, University of California, San Diego , ⁵Broad Institute of Harvard and MIT, ⁶Department of Medicine, Brigham and Women's Hospital, ⁷Electrical Engineering and Computer Science and David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, ⁸Howard Hughes Medical Institute

The goal of these experiments is to generate quantitative time-course data on the growth and gene expression dynamics of attenuated S. typhimurium bacterial colonies growing inside tumors. This video covers tumor cell preparation and implantation, bacteria preparation and injection, whole-animal luminescence imaging, tumor excision, and bacterial colony counting.

JoVE Core

Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation

Emily A. Brown^1,2, Steven C. Neier^3,4, Claudia Neuhauser⁵, Adam G. Schrum^6,7,8, Stephen E.P. Smith^1,2,9

¹Center for Integrative Brain Research, Seattle Children's Research Institute, ²Graduate Program in Neuroscience, University of Washington, ³Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Department of Medicine, Harvard Medical School, ⁴Broad Institute of Harvard and MIT, ⁵Department of Mathematics, University of Houston, ⁶Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, ⁷Department of Surgery, School of Medicine, University of Missouri, ⁸Department Bioengineering, College of Engineering, University of Missouri, ⁹Department of Pediatrics, University of Washington

Quantitative Multiplex Immunoprecipitation (QMI) uses flow cytometry for sensitive detection of differences in the abundance of targeted protein-protein interactions between two samples. QMI can be performed using a small amount of biomaterial, does not require genetically engineered tags, and can be adapted for any previously defined protein interaction network.