Screening of eGFP expressing transgenic mice

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ashwinraut
ashwinraut's picture
Screening of eGFP expressing transgenic mice

I am going to produce a population of transgenic mice using lentiviral transduction methods, using sperm mediated transgenesis where eGFP will be used as reporter. Only 10% population is expected to be transgenic producing eGFP and the rest will be negative. Kindly suggest me a easy method to screening the eGFP producing transgenic mice from mixed population of eGFP expressing and non expressing mice population.

atcg
atcg's picture
Ashwinraut - perhaps this

Ashwinraut - perhaps this article would help. A group of scientists did a study on the cellular distribution of enhanced green flourescent protein (EGFP)in the retina. They used transgenic and nontransgenic litter mates - so the article may describe how they sorted the mixed population of eGFP expressing mice. The link below will take you to the Investigative Opthamology and Visual Science Article:

" Absence of Functional and Structural Abnormalities Associated with Expression of EGFP in the Retina"

http://www.iovs.org/cgi/content/abstract/45/1/15

Written by: May Nour,1 Alexander B. Quiambao,2 Muayyad R. AlUbaidi,1,2 and Muna I. Naash1,2
1From the Oklahoma Center for Neuroscience, and the 2Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma.

PURPOSE. The present study was undertaken to evaluate the effect of uniform EGFP expression on retinal morphology and function.

Hope this helps : )

ashwinraut
ashwinraut's picture
Thank you for the reference.

Thank you for the reference. I had gone through the article but procedure described is too tedious and specifically mentioned for retina so of little help.
Ashwin Raut

Omai
Omai's picture
I thought it was important to

I thought it was important to bring up this older post to share with the scientist solutions community some recent advances in fluorescent protein carrying mouse strains. Jackson now provides over 16 different mouse strains containg fluorescent reporters. They come in multiple excitation and emission wave lengths.

Here is a little blurb from the website on visualization stratagies and some links to companies who provide this equipment (which is the focus of this post).

"There are numerous ways to visualize the bioluminescence of fluorescent proteins. One way to detect the fluorescence is with a hand-held UV light (~365 nm). However, the optimum excitation wavelength for GFP is 488 nm. CFP will fluoresce cyan when exposed to a 433 nm light source, and YFP will fluoresce yellowish-green when exposed to a 513 nm light source. It is best to use the light source that corresponds to the optimum excitation wavelength for each specific fluorescent protein.
Andras Nagy of The Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Ontario, has a Web site which discusses the use of fluorescent constructs in transgenic mice (http://www.mshri.on.ca/nagy/). Wallace Marshall at Yale University has also created a Web resource for researchers interested in working with GFP (http://pantheon.cis.yale.edu/~wfm5/gfp_gateway.html).
The "Mouse Models Expressing Fluorescent Proteins or lacZ" list, available on the JAX® Mice Web site at http://jaxmice.jax.org/html/pricelist/fluorescent.lacz.pdf, describes in further detail models which express fluorescent proteins.
Sources for equipment for visualization of fluorescent proteins are provided below.  This list is not comprehensive, and is provided for information purposes only and does not represent endorsement by The Jackson Laboratory."
BLS Ltd.
http://www.bls-ltd.com/
Chroma Technology Corp.
http://www.chroma.com
Lightools Research
http://www.lightools.com
UVP, Inc.
http://www.uvp.com
Finally, here is a picture of some GFP mice, just for fun from this link:

Omai