Optogenetic dephosphorylation of phosphatidylinositol 4,5 bisphosphate in Xenopus laevis oocytes.

Here, we present a protocol for optogenetic dephosphorylation of the phosphoinositide PI(4,5)P ₂ at the plasma membrane of Xenopus laevis oocytes. We first describe the co-injection of oocytes with cRNAs encoding (1) a light-activated PI(4,5)P ₂ 5-phosphatase fusion protein, (2) its dimerization partner fused to the plasma membrane, and (3) the potassium channel reporter for PI(4,5)P ₂ dephosphorylation. We then detail blue light illumination to induce PI(4,5)P ₂ dephosphorylation, combined with simultaneous two-electrode voltage clamp electrophysiological recording to assess potassium channel current responses. For complete details on the use and execution of this protocol, please refer to Xu et al. (2022). ¹ .
Competing Interests: Declaration of interests The authors declare no competing interests.
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