Phosphoinositide (PI) concentrations in cell membranes play an important role in many cellular processes. PI phosphatases are crucial for the regulation of these concentrations. Among these phosphatases, a unique family, the voltage-sensitive phosphatases (VSPs), provides a new paradigm for control of enzymatic activity. Recently, we showed that voltage sensitivity can be conferred to the phosphatase PTEN by fusing the voltage sensor domain (VSD) of Ci-VSP, the prototypical VSP from Ciona intestinalis, to the catalytic domain (CD) of PTEN. Here we extend this work and report on a chimera created by fusing Ci-VSP's VSD to the CD of the PI-phosphatase TPIPa. We demonstrate that the Ci-VSP/TPIPa chimera exhibits voltage dependent enzymatic activity. The substrate specificity of Ci-VSP/TPIPa is analyzed using genetically encoded PI-specific fluorescence labeled probes, the membrane binding of which is assessed using total internal reflection fluorescence (TIRF) microscopy. We find that upon depolarization, the membrane binding of the PI(3,4,5)P₃ specific Btk-PH-GFP and the PI(4,5)P₂ specific PLCd₁-PH-GFP probes decreases, when co-expressed with Ci-VSP/TPIPa. In contrast, membrane binding of the PI(3,4)P₂ specific TAPP1-PH-GFP and the PI(4)P specific OSBP-PH-GFP probes increases. These findings indicate that the catalytic domain of TPIPa functions as a PI(4,5)P₂/PI(3,4,5)P₃-5'-phosphatase when under control of a VSD. These data will help to further the understanding of the mechanism by which voltage control is exerted in VSPs. Additionally, our experiments demonstrate the usefulness of engineered VSPs as a novel tool for the analysis of PI-phosphatases in-vivo, an obligatory complement to in-vitro characterization. Supported by Deutsche Forschungsgemeinschaft (SFB593 TPA12 to D.O.)