Evidence map›Paper›PMID 31879907›Full record

ArticleMethods in molecular biology (Clifton, N.J.)2020

Optogenetic Control of Microtubule Dynamics.

Jeffrey van Haren, Lauren S Adachi, Torsten Wittmann

Abstract read
In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed.

  1. Article
  2. Article
  3. Article
  4. Review
  5. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

3 authors.

Jeffrey van HarenDepartment of Cell and Tissue Biology, University of California San Francisco, San Francisco, CA, USA.
Lauren S AdachiDepartment of Cell and Tissue Biology, University of California San Francisco, San Francisco, CA, USA.
Torsten WittmannDepartment of Cell and Tissue Biology, University of California San Francisco, San Francisco, CA, USA. Torsten.Wittmann@ucsf.edu.

Funding

Probing Microtubule Function in Neuronal DevelopmentR01NS107480 · NINDS · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI WITTMANN, TORSTEN · 2018 to 2022
$2.0M
Light-activated proteolysis as a tool to analyze intracellular protein functionR01GM094819 · NIGMS · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI WITTMANN, TORSTEN · 2010 to 2013
$1.2M
Spinning disk confocal / FRAP microscope for quantitative live cell imagingS10RR026758 · NCRR · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI WITTMANN, TORSTEN · 2010 to 2010
$500k
Dissecting protein activities by reversible photo-inactivationR21CA224194 · NCI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI WITTMANN, TORSTEN · 2017 to 2018
$379k
NCI NIH HHS R21 CA224194NCRR NIH HHS S10 RR026758NIGMS NIH HHS R01 GM094819NINDS NIH HHS R01 NS107480
6 · The paper itself

Abstract

Light can be controlled with high spatial and temporal accuracy. Therefore, optogenetics is an attractive experimental approach to modulate intracellular cytoskeleton dynamics at much faster timescales than by genetic modification. For example, in mammalian cells, microtubules (MTs) grow tens of micrometers per minute and many intracellular MT functions are mediated by a complex of +TIP proteins that dynamically associate with growing MT plus ends. EB1 is a central component of this +TIP protein network, and we recently developed a photo-inactivated π-EB1 by inserting a blue light-sensitive LOV2/Zdk1 module between the EB1 MT-binding domain and the +TIP adaptor domain. Blue light-induced π-EB1 photodissociation results in disassembly of the +TIP complex and strongly attenuates MT growth in mammalian cells.In this chapter, we discuss theoretical and practical aspects of how to perform high-resolution live-cell microscopy in combination with π-EB1 photodissociation. However, these techniques are broadly applicable to other LOV2-based and likely other blue light-sensitive optogenetics. In addition to being a tool to investigate +TIP functions acutely and with subcellular resolution, because of its dramatic and rapid change in intracellular localization, π-EB1 can serve as a powerful tool to test and characterize optogenetic illumination setups. We describe protocols on how to achieve micrometer-scale intracellular control of π-EB1 activity using patterned illumination, and we introduce a do-it-yourself LED cube design compatible with transmitted light microscopy in multiwell plates.

Indexed as

Microscopy, FluorescenceOptogeneticsAnimalsCell LineHumansMicrotubule-Associated ProteinsMicrotubulesProtein BindingProtein Interaction Domains and MotifsTime-Lapse ImagingMicrotubule-Associated ProteinsEB1Live-cell microscopyLOV2LOVTRAPMicrotubulesOptogeneticsPhotodissociation+TIPZdk1π-EB1

Identifiers

PMID31879907
PMCPMC7278073

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Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.