Naslov
Optogenetic reversible knocksideways, laser ablation, and photoactivation on the mitotic spindle in human cells

Autori
Milas, Ana ; Jagrić, Mihaela ; Martinčić, Jelena ; Tolić, Iva M.

Vrsta, podvrsta i kategorija rada
Poglavlja u knjigama, znanstveni

Knjiga
Methods in Cell Biology

Urednik/ci
Maiato, Helder ; Schuh, Melina

Izdavač
Elsevier

Godina
2018

Raspon stranica
191-215

ISBN
978-0-12-814142-7

ISSN
0091-679X

Ključne riječi
Mitosis ; Mitotic spindle ; Optogenetics ; Light-induced dimerization ; Photosensitive domain ; Laser ablation ; Laser microsurgery ; Laser cutting ; Photoactivation ; Kinetochore fibers

Sažetak
At the onset of mitosis, cells assemble the mitotic spindle, a dynamic micromachine made of microtubules and associated proteins. Although most of these proteins have been identified, it is still unknown how their collective behavior drives spindle formation and function. Over the last decade, RNA interference has been the main tool for revealing the role of spindle proteins. However, the effects of this method are evident only after a longer time period, leading to difficulties in the interpretation of phenotypes. Optogenetics is a novel technology that enables fast, reversible and precise control of protein activity by utilization of light. In this chapter, we present an optogenetic knocksideways method for rapid and reversible translocation of proteins from the mitotic spindle to mitochondria using blue light. Furthermore, we discuss other optical approaches, such as laser ablation of microtubule bundles in the spindle and creation of reference marks on the bundles by photoactivation of photoactivatable GFP. Finally, we show how different optical perturbations can be combined in order to acquire deeper understanding of the mechanics of mitosis.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA