Reporter Core | Chemistry Core | High Throughput Core

Reporter Core
Scientific Co-Director - Samuel Achilefu
Scientific Co-Director - David Piwnica-Worms
Contact - Julie Prior

Mission Statement

The Molecular Imaging Reporter Core (MIRC) is a central facility that not only develops and generates new reporter constructs, cell lines and genetically engineered animals, but also provides expertise, materials and collaborative assistance for design and execution of the biological aspects of molecular imaging. The MIRC serves investigators with a wide range of resources and experience in molecular biology, mammalian cell culture, and small animal experimentation. One of the most important service activities of this core is dissemination of our newly developed molecular imaging reagents, reporter cell lines and genetically-encoded reporters to investigators within our institution, to other imaging institutions and to cancer biology investigators throughout the world.

Functionality Description

Discovery Research

The MIRC is also highly active in discovery based research, creating new constructs and generating transgenic reporter animals.

Service and Expertise

While these web-based catalogues provide an archive of information useful for experienced investigators, the MIRC also provides assistance to investigators who have limited experience or resources by advice or direct help with experimental design and execution. The Core can clone and screen reporters and generate cell lines and transgenic or knock-in animals. Furthermore, MIRC can characterize reporter gene expression and molecular function using either radio-labeled or fluorescently tagged probes (together with the Chemistry Core) or through bioluminescence assays in vitro. Additionally, the MIRC assists in planning and executing xenograft and transgenic studies in vivo (see Imaging Resources).

Plasmid Repository

The plasmid repository contains general information about plasmids (e.g. promoter, insert) as well as cloning strategies. Bacterial and/or mammalian selection markers are described. Descriptions of the vector backbone and a circular map are accessible. Furthermore, the DNA sequence file can be retrieved which allows manipulation using any standard DNA software.

The MIRC serves as a central clearinghouse for molecular imaging reagents and protocols related to biological aspects of molecular imaging. These materials include reporters for genetic transduction (plasmids and viruses) to establish cell lines that stably express imaging constructs for in vitro assays or xenograft production or to create transgenic reporter mice. All information is catalogued in a repository database.

Viral Constructs
Map
Sequence
Reference
Request
FUW-FLG
FUW-CBR-mCherry
 
FUW-CBG-GFP
 
Split-Fluc Reporters
Map
Sequence
Reference
Request
FRB-Nluc
Cluc-FKBP
IκBα Reporters
Map
Sequence
Reference
Request
CMV→IκBα-Fluc
CMV→Fluc
(control for CMV→IκBα-Fluc)
κBx5→IκBα-Fluc
κBx5→Fluc
(control for κBx5→IκBα-Fluc)
β-catenin Reporters
Map
Sequence
Reference
Request
CMV→β-cat-Fluc
CMV→Fluc
(control for CMV→β-cat-Fluc)
BRET Reporters
Map
Sequence
Reference
Request
CBG68-DEVD-tdTomato
CBG68-GGSS-tdTomato
Miscellaneous
Map
Sequence
Reference
Request
CMV→Ubx4-Fluc
Gal4→Fluc
MDR1-Fluc

For a full list of plasmids available from Addgene, click here

Cell Lines

A number of reporter cell lines are available. For each specific cell line, source laboratory, growth media and culturing conditions, use in animal models, and other relevant information are contained within accompanying data sheets.

Name
Cell Type
Reference
4T1 murine breast cancer (balb/c)
4T1-GFP-FL murine breast cancer (balb/c)
B16-FL murine melanoma (BL/6)
LLC G/L murine lung cancer (BL/6)  
SKOV3-Luc eYFP human ovarian cancer
U87-GL human GBM cancer
U266 human multiple myeloma  
OPM2 human multiple myeloma  
5TGM1-GFP murine multiple myeloma
MM1.S human multiple myeloma  
MM1.S-5(CBR-GFP) human multiple myeloma  
MM1.S-9(CBR-GFP) human multiple myeloma  

Transgenic Animals

Transgenic animals are created with the coordinated help of two other core facilities at Washington University, the Mouse Embryonic Stem Cell Core or the Mouse Genetics Core. Both facilities have the required expertise and infrastructure for these important steps in generating transgenic animals and provide high quality service to the WU research community.

Cryopreserved sperm is available for the following transgenic mouse lines:

Name
Background Strain
Reference
Gal4→Fluc Mouse
FVB
ROSA26→CBRlacZ Mouse
albino C57BL/6
 
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