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Engineering oscillating microtubule bundles.

Methods in enzymology (2013-03-19)
Timothy Sanchez, Zvonimir Dogic
ABSTRACT

From motility of simple protists to determining the handedness of complex vertebrates, highly conserved eukaryotic cilia and flagella are essential for the reproduction and survival of many biological organisms. Despite extensive studies, the exact mechanism by which individual components coordinate their activity to produce ciliary beating patterns remains unknown. We describe a novel approach toward studying ciliary beating. Instead of deconstructing a fully functional organelle from the top-down, we describe a process by which synthetic cilia-like structures are assembled from the bottom-up and we present methods for engineering such structures. We demonstrate how simple mixtures of microtubules, kinesin clusters, and a bundling agent assemble into structures that produce spontaneous oscillations, suggesting that self-organized beating may be a generic feature of internally driven bundles. Synthetic cilia-like structures can be assembled at high density, leading to synchronization and metachronal traveling waves, reminiscent of the waves seen in biological ciliary fields.

MATERIALS
Product Number
Brand
Product Description

Supelco
Biotin, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
Biotin, tested according to Ph. Eur.
Sigma-Aldrich
Biotin, ≥99.0% (T)
Sigma-Aldrich
Biotin, meets USP testing specifications
Sigma-Aldrich
Biotin, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Biotin, European Pharmacopoeia (EP) Reference Standard
Supelco
Biotin, Pharmaceutical Secondary Standard; Certified Reference Material