Measuring Growth and Gene Expression Dynamics of Tumor-Targeted S. Typhimurium Bacteria
Name
Danino-2013-Measuring growth and.pdf
Size
290.22 KB
Format
Adobe PDF
Checksum (MD5)
9f710a61e79687a68fab220f8b240b1d
Author(s)
Danino, Tal
Prindle, Arthur
Hasty, Jeff
Bhatia, Sangeeta N.
Bhatia, Sangeeta N
Date Issued
July 2013
Journal
Journal of Visualized Experiments
Publisher
MyJoVE Corporation
Citation
Danino, Tal, Arthur Prindle, Jeff Hasty, and Sangeeta Bhatia. “Measuring Growth and Gene Expression Dynamics of Tumor-Targeted S. Typhimurium Bacteria.” JoVE no. 77 (2013).
Version
Final published version
Abstract
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.
We generated model xenograft tumors in mice by subcutaneous injection of a human ovarian cancer cell line, OVCAR-8 (NCI DCTD Tumor Repository, Frederick, MD).
We transformed attenuated strains of S. typhimurium bacteria (ELH430:SL1344 phoPQ- 1) with a constitutively expressed luciferase (luxCDABE) plasmid for visualization2. These strains specifically colonize tumors while remaining essentially non-virulent to the mouse1.
Once measurable tumors were established, bacteria were injected intravenously via the tail vein with varying dosage. Tumor-localized, bacterial gene expression was monitored in real time over the course of 60 hours using an in vivo imaging system (IVIS). At each time point, tumors were excised, homogenized, and plated to quantitate bacterial colonies for correlation with gene expression data.
Together, this data yields a quantitative measure of the in vivo growth and gene expression dynamics of bacteria growing inside tumors.
MIT Department
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Koch Institute for Integrative Cancer Research at MIT. Laboratory for Multiscale Regenerative Technologies
Koch Institute for Integrative Cancer Research at MIT
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.3791/50540
