Tattletales and T-Bow Update 20140602Mon
Please see also http://works.bepress.com/gmcnamara/26
Tattletales: multiplex fluorescent protein biosensors by spatial localization with TALE-FPs, Cas9-FPs, ZF-FPs, LacI-FPs, TetR-FPs, etc.
T-Bow: Rainbow T-cells and Tumor cells (and ES cells, iPS cells, other cells and organisms). You can think of this as "Brainbow meets TALENs/Cas9/ZFNs/other DNA sequence specific binding proteins".
If not familiar with Brainbow, see
If not familiar with TALENs, Cas9, etc, see
Big idea: localizing fluorescent proteins - and/or Nano-Lanterns (Take Nagai) - to tandem repeat arrays - is a great way to improve signal to noise ratio compared to the usual "overexpressed FP smog" AND to multiplex saeveral biosensors and other reporters (ex: promoter bashing).
See Figure 4A of Robinett et al 1996 JCB
(freely available) for first example of "single locus" (256 element LacO tandem repeat array) using LacI-nls-GFP fusion protein (512 FPs at array, smog from overexpression).
In Nov 2013-Feb 2014 several labs have published TALE-FPs and Cas9-FPs, see the reference list in the enclosed PDF(s). Several mention patent applications: too bad for their bank accounts that Robinett et al's 1996 prior art, and my public disclosure in October 2013
makes TALE-FPs and Cas9-FPs obvious (my public disclosure being inspired by Robinett et al's publication). I suspect some "validated" FP-fusion cell lines will be patented - very likely can be duplicated in any biomedical research lab for a lot less than the licensing fees desired by the tech transfer folks.
George McNamara. "Tattletales and T-Bow Update 20140602M" 2014
Available at: http://works.bepress.com/gmcnamara/42
"42" was originally posted 20150505 (May 5, 2014). On 20140602M (June 20, 2014, Monday) I updated a couple of files in the ZIP download. The FP Biosensors summary table is now at 130 entries. A presentation has been added that includes "Binary Tattletales Summary" (penultimate slide in the pdf file dated today).
- Rainbow T-cells and Tumor Cells,
- Promoter bashing,
- Fluorescent Protein Biosensors,
Available at: http://works.bepress.com/gmcnamara/42/