C04722: Spatiotemporal FRET Reporter for PhosphoinositidesNovelty:
This technology is a novel reporter molecule with the capability of revealing very specific spatial and temporal information for production and degradation of phosphatidylinositols (PI) PIP(3,4)2 and PIP3. Phosphatidylinositols are a diverse family of lipid second messengers with important roles in various cellular processes.
Value Proposition:
PIs are important second messengers that are responsible for the recruitment of protein kinase B, or Akt, to the plasma membrane. Akt plays many important roles in various cellular processes including cell proliferation, differentiation, survival and tumorigenesis. Due to the diversity of function that these PIs are involved with, cellular control of their spatial and temporal dynamics is crucial, yet there is no current reliable method available for measuring this. Additionally, current imaging methods for PIs are wrought with issues ranging from limitations of resolution to artifacts. This technology solves these issues through a novel emission ratiometric reporter called InPAkt and its use with Fluorescent Resonance Energy Transfer (FRET) technology to monitor the production and degradation of PIs with high temporal and spatial resolution. Additional advantages include:
• Targeted reporter to subcellular organelles for monitoring specific pools of PIs
• Able to undergo conformational change without membrane anchoring
• Small amounts of tissue or cells required
• Dual readout of FRET change and translocation of its untargeted form
• Eliminates any artifacts cause by cell shape change
Technical Details:
Johns Hopkins researchers have developed a novel emission ratiometric reporter, InPAkt, as a research tool for monitoring the production and degradation of PIP(3,4)2 and PIP3. The reporter molecule comprises of a pair of fluorescent proteins linked by a region containing a phosphoinositide binding domain, the pleckstrin homology domain of Akt, and a pseudoligand of acidic amino acids. Upon binding of PIP(3,4)2 or PIP3 , the reporter will undergo a conformational change resulting in a different FRET emission ratio.
Looking for Partners:
To develop and commercialize this innovation as a novel research tool for increased understanding of phosphatidylinositols.
Stage of Development:
Research tool published
Data Availability:
Under CDA/ NDA
Publications/Associated Cases:
Proc Natl Acad Sci U S A. 2005 Oct 18;102(42):15081-6
