Light-Powered Reactivation of Flagella and Contraction of Microtubule Networks: Toward Building an Artificial Cell.

Raheel Ahmad,Eberhard Bodenschatz,Isabella Guido,Albert Bae,Vahid Nasirimarekani,Tanja Vidaković-Koch,Kai Sundmacher,Yu-Jung Su,Samira Goli Pozveh,Azam Gholami,Christin Kleineberg

doi:10.1021/acssynbio.1c00071

Abstract

Artificial systems capable of self-sustained movement with self-sufficient energy are of high interest with respect to the development of many challenging applications, including medical treatments, but also technical applications. The bottom-up assembly of such systems in the context of synthetic biology is still a challenging task. In this work, we demonstrate the biocompatibility and efficiency of an artificial light-driven energy module and a motility functional unit by integrating light-switchable photosynthetic vesicles with demembranated flagella. The flagellar propulsion is coupled to the beating frequency, and dynamic ATP synthesis in response to illumination allows us to control beating frequency of flagella in a light-dependent manner. In addition, we verified the functionality of light-powered synthetic vesicles in in vitro motility assays by encapsulating microtubules assembled with force-generating kinesin-1 motors and the energy module to investigate the dynamics of a contractile filamentous network in cell-like compartments by optical stimulation. Integration of this photosynthetic system with various biological building blocks such as cytoskeletal filaments and molecular motors may contribute to the bottom-up synthesis of artificial cells that are able to undergo motor-driven morphological deformations and exhibit directional motion in a light-controllable fashion.

Highlights

Artificial systems capable of self-sustained movement with self-sufficient energy are of high interest with respect to the development of many challenging applications, including medical treatments, and technical applications
The reconstitution of each of these processes is quite complex, so the bottom-up synthetic biology initially focuses on subsystems, which are hand-tailored to partially or entirely mimic some of the essential life processes.[1−4] Later on, these functional modules can be combined to build higher hierarchical structures
The regular beating pattern of cilia and flagella propels microorganisms such as spermatozoa, or unicellular organisms such as Paramecium and green algae Chlamydomonas reinhardtii.[11−14] Cilia and flagella are highly conserved organelles composed of a microtubule-based structure called axoneme, which is covered by a plasma membrane.[19,20]

Summary

Introduction

Artificial systems capable of self-sustained movement with self-sufficient energy are of high interest with respect to the development of many challenging applications, including medical treatments, and technical applications. We verified the functionality of light-powered synthetic vesicles in in vitro motility assays by encapsulating microtubules assembled with force-generating kinesin-1 motors and the energy module to investigate the dynamics of a contractile filamentous network in cell-like compartments by optical stimulation. Integration of this photosynthetic system with various biological building blocks such as cytoskeletal filaments and molecular motors may contribute to the bottom-up synthesis of artificial cells that are able to undergo motor-driven morphological deformations and exhibit directional motion in a light-controllable fashion. The ultimate challenging goal in the growing field of synthetic biology is to build an artificial cell with a selfsufficient energy conversion system to autonomously power the activity of molecular motors

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