Extended Data Fig. 2: Rational behind the design of the iDePP system.
From: Inducible depletion of PI(4,5)P2 by the synthetic iDePP system in Arabidopsis
a, Schematic drawing depicting constructs used to genetically-induced PI(4,5)P2 pool modification at the PM. After dex treatment, MAP-mCh-dOCRL is anchored at the plasma membrane and dephosphorylates PI(4,5)P2, causing the PI(4,5)P2 biosensor to be released into the cytosplasm. b, Controls used in this study includes 3xmCh and a catalytic dead version of dOCRL. After dex treatment, both MAP-3xmCh and MAP-mCh-dOCRLdead are anchored at the PM, but do not affect the pool of PI(4,5)P2. The PI(4,5)P2 biosensor is found at the PM. To build the iDePP system, we had to engineer a synthetic enzyme, which consists of the isolated catalytic domain of a 5-phosphatase (which if expressed on its own would be soluble in the cytosol and likely inactive) and then target this module to the plasma membrane, for specific depletion of PI(4,5)P2). Details can be found in the Methods section.
