Evidence map›Paper›PMID 31544194›Full record

ArticleLab on a chip2019

Light-inducible activation of cell cycle progression in Xenopus egg extracts under microfluidic confinement.

Jitender Bisht, Paige LeValley, Benjamin Noren, Ralph McBride, Prathamesh Kharkar, April Kloxin, Jesse Gatlin, John Oakey

Open access · greenAbstract read
In one paragraph

Article in Lab on a chip, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
0.2field-weighted citation impact, top 49% of its field
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

4 citing papers in PubMed, 7 citations in OpenAlex.

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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

8 authors at 2 institutions in 2 countries.

Jitender BishtDepartment of Molecular Biology, University of Wyoming, Laramie, WY 82071, USA. jgatlin@uwyo.edu.
Paige LeValley
Benjamin Noren
Ralph McBride
Prathamesh Kharkar
April Kloxin
Jesse Gatlin
John Oakey
University of Wyoming · USNewark Hospital · GB

Funding

Wyoming INBRE Phase 4- Equipment Supplement for x-ray diffractometer for Center for Advanced Scientific InstrumentationP20GM103432 · NIGMS · UNIVERSITY OF WYOMING · PI Robert Scott Seville · 2012 to 2026
$56.8M
This renovation project will create over 1455 sq. ft. of state- of-the-art reseaP20GM104316 · NIGMS · UNIVERSITY OF DELAWARE · PI FOX, JOSEPH M · 2014 to 2024
$26.8M
Pilot Research Subproject ProgramP30GM110758 · NIGMS · UNIVERSITY OF DELAWARE · PI BEEBE, THOMAS PAUL · 2014 to 2018
$5.8M
Integration of Xenopus extract and microfluidics to study organelle size scalingR01GM113028 · NIGMS · UNIVERSITY OF WYOMING · PI LEVY, DANIEL LEON · 2015 to 2019
$1.3M
Spindle Assembly and Scaling via Microfluidic Encapsulation of Xenopus NucleiR15GM101636 · NIGMS · UNIVERSITY OF WYOMING · PI OAKEY, JOHN S · 2012 to 2012
$313k
NIGMS NIH HHS P20 GM103432NIGMS NIH HHS P20 GM104316NIGMS NIH HHS P30 GM110758NIGMS NIH HHS R01 GM113028NIGMS NIH HHS R15 GM101636
6 · The paper itself

Abstract

Cell-free Xenopus egg extract is a widely used and biochemically tractable model system that allows recapitulation and elucidation of fundamental cellular processes. Recently, the introduction of microfluidic extract manipulation has enabled compartmentalization of bulk extract and a newfound ability to study organelles on length scales that recapitulate key features of cellular morphology. While the microfluidic confinement of extracts has produced a compelling platform for the in vitro study of cell processes at physiologically-relevant length scales, it also imposes experimental limitations by restricting dynamic control over extract properties. Here, we introduce photodegradable polyethylene glycol (PEG) hydrogels as a vehicle to passively and selectively manipulate extract composition through the release of proteins encapsulated within the hydrogel matrix. Photopatterned PEG hydrogels, passive to both extract and encapsulated proteins, serve as protein depots within microfluidic channels, which are subsequently flooded with extract. Illumination by ultraviolet light (UV) degrades the hydrogel structures and releases encapsulated protein. We show that an engineered fluorescent protein with a nuclear localization signal (GST-GFP-NLS) retains its ability to localize within nearby nuclei following UV-induced release from hydrogel structures. When diffusion is considered, the kinetics of nuclear accumulation are similar to those in experiments utilizing conventional, bulk fluid handling. Similarly, the release of recombinant cyclin B Δ90, a mutant form of the master cell cycle regulator cyclin B which lacks the canonical destruction box, was able to induce the expected cell cycle transition from interphase to mitosis. This transition was confirmed by the observation of nuclear envelope breakdown (NEBD), a phenomenological hallmark of mitosis, and the induction of mitosis-specific biochemical markers. This approach to extract manipulation presents a versatile and customizable route to regulating the spatial and temporal dynamics of cellular events in microfluidically confined cell-free extracts.

Indexed as

MitosisUltraviolet RaysAnimalsCell NucleusCyclin BHydrogelsMicrofluidicsOocytesPolyethylene GlycolsRecombinant ProteinsXenopus laevisCyclin BHydrogelsPolyethylene GlycolsRecombinant Proteins

Identifiers

PMID31544194
PMCPMC7819639
OpenAlexW2971392101

What OpenQuestion holds

Textmetadata
LicenceTDM
Read underepoch 390

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.