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. Author manuscript; available in PMC: 2016 Jun 1.
Published in final edited form as: Biochim Biophys Acta. 2014 Dec 13;1851(6):736–745. doi: 10.1016/j.bbalip.2014.12.008

Fig. 3.

Fig. 3

Genetic tools used to acutely manipulate PI levels in live cells. A. Electrogenetic manipulation is based on the ectopic expression of a voltage-sensing domain (VSP) fused to a PI-metabolizing enzyme (PIM). This fusion protein is inserted into the plasma membrane (PM) and enzymatic activity is stimulated by plasma membrane depolarization. B. Chemical genetic manipulation requires the expression of FRB anchored to a target membrane (TM) and FKBP12 fused to a PI-metabolizing enzyme (PIM) or vice versa. Addition of rapamycin (or a rapalog) promotes FRB-FKBP dimerization and brings the enzyme to the target membrane where it will act on its PI(s) targets. C. Optogenetic manipulation is based on blue light-induced dimerization between a membrane-anchored protein (CIBN) and its binding partner (CRY2) fused to a PI-metabolizing enzyme (PIM). PhyB and PIF6 can be used as alternative light-sensitive dimerization pairs.