Author ORCID Identifier

https://orcid.org/0009-0001-3350-2176

Date of Award

5-12-2026

Document Type

Thesis (Undergraduate)

Department

Biological Sciences

First Advisor

Bing He

Second Advisor

Patrick Dolph

Third Advisor

Wei-Lih Lee

Abstract

Intracellular trafficking plays a central role in tissue morphogenesis by enabling cell signaling involved in spatial patterning, regulating tissue architecture, and mediating feedback responses to mechanical forces. Among trafficking regulators, the small GTPase Rab11 is a key regulator of recycling endosome function and polarized membrane delivery. It is involved in both exocytic and endocytic trafficking pathways, and it is transported by multiple motor proteins depending on the context. Conventional genetic perturbations of trafficking regulators during development are often difficult to interpret, as these proteins are essential to various aspects of morphogenesis and often have distinct, stage-specific functions. To address this limitation, we developed and validated optogenetic control of Rab11-mediated trafficking using Opto-Rab11-kinesin, a CRY2-CIBN-based tool that enables blue-light-induced recruitment of the kinesin motor protein to Rab11 compartments. We characterized the effects of Opto-Rab11-kinesin activation on Rab11 localization across multiple developmental contexts in live Drosophila embryos and show that this approach effectively recruits Rab11 compartments to the plus end of microtubules. We further applied this tool to investigate the role of Rab11 during apical constriction, a conserved morphogenetic process driven by actin- and myosin-mediated contractility. We found that acute Opto-Rab11-kinesin activation resulted in altered myosin organization, unproductive myosin contractions and failed mesoderm invagination during Drosophila gastrulation. These findings establish Opto-Rab11-kinesin as a powerful tool for studying stage-specific functions of Rab11-mediated trafficking and provide new insights into the role of Rab11 in regulating the apical actomyosin network during morphogenesis.

Available for download on Friday, July 16, 2027

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