Growth cone advance requires EB1 as revealed by genomic replacement with a light-sensitive variant: One-step engineering of photosensitive EB1 in hiPSCs and derived neurons

Alessandro Dema, Rabab A Charafeddine, Shima Rahgozar, Jeffrey van Haren, Torsten Wittmann*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Scopus)
31 Downloads (Pure)

Abstract

A challenge in analyzing dynamic intracellular cell biological processes is the dearth of methodologies that are sufficiently fast and specific to perturb intracellular protein activities. We previously developed a light-sensitive variant of the microtubule plus end tracking protein EB1 by inserting a blue light-controlled protein dimerization module between functional domains. Here, we describe an advanced method to replace endogenous EB1 with this light-sensitive variant in a single genome editing step, thereby enabling this approach in human induced pluripotent stem cells (hiPSCs) and hiPSC-derived neurons. We demonstrate that acute and local optogenetic EB1 inactivation in developing cortical neurons induces microtubule depolymerization in the growth cone periphery and subsequent neurite retraction. In addition, advancing growth cones are repelled from areas of blue light exposure. These phenotypes were independent of the neuronal EB1 homolog EB3, revealing a direct dynamic role of EB1-mediated microtubule plus end interactions in neuron morphogenesis and neurite guidance.

Original languageEnglish
Article numbere84143
JournaleLife
Volume12
Early online date30 Jan 2023
DOIs
Publication statusPublished - Jan 2023

Bibliographical note

This work was supported by National Institutes of Health grants R21 CA224194, R01 NS107480, S10 RR026758 and S10 OD028611 to T.W.

© 2023, Dema et al.

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