Evidence map›Paper›PMID 34425213›Full record

ArticleFungal genetics and biology : FG & B2021

Quantitative single molecule RNA-FISH and RNase-free cell wall digestion in Neurospora crassa.

Bradley M Bartholomai, Amy S Gladfelter, Jennifer J Loros, Jay C Dunlap

Open access · greenAbstract read
In one paragraph

Article in Fungal genetics and biology : FG & B, 2021. 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 47% 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, 4 citations in OpenAlex.

  1. Article
  2. Article
  3. PRD-2 mediates clock-regulated perinuclear localization of clock gene RNAs within the circadian cycle ofProceedings of the National Academy of Sciences of the United States of America · 2022
    Article
  4. Advances and Challenges in FluorescenceFrontiers in microbiology · 2022
    Article
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

4 authors at 2 institutions in 1 country.

Bradley M BartholomaiGeisel School of Medicine at Dartmouth, Department of Molecular and Systems Biology, Hanover, NH, USA.
Amy S GladfelterUniversity of North Carolina, Department of Biology, Chapel Hill, NC, USA.
Jennifer J LorosGeisel School of Medicine at Dartmouth, Department of Biochemistry and Cell Biology, Hanover, NH, USA.
Jay C DunlapGeisel School of Medicine at Dartmouth, Department of Molecular and Systems Biology, Hanover, NH, USA. Electronic address: jay.c.dunlap@dartmouth.edu.
Dartmouth College · USUniversity of North Carolina at Chapel Hill · US

Funding

Understanding the role of RNA-binding protein mutations in cancerP20GM113132 · NIGMS · DARTMOUTH COLLEGE · PI MADDEN, DEAN R · 2016 to 2025
$25.9M
Genetic and Molecular Dissection of the Neurospora ClockR35GM118021 · NIGMS · DARTMOUTH COLLEGE · PI Jay C. Dunlap · 2016 to 2026
$7.3M
Molecular and Cellular Biology at DartmouthT32GM008704 · NIGMS · DARTMOUTH COLLEGE · PI COMPTON, DUANE A. · 1998 to 2018
$6.8M
Identification and Analysis of Circadian Clock-Controlled GenesR35GM118022 · NIGMS · DARTMOUTH COLLEGE · PI LOROS, JENNIFER J. · 2016 to 2021
$3.7M
NIGMS NIH HHS P20 GM113132NIGMS NIH HHS R35 GM118021NIGMS NIH HHS R35 GM118022NIGMS NIH HHS T32 GM008704
6 · The paper itself

Abstract

Single molecule RNA-FISH (smFISH) is a valuable tool for analysis of mRNA spatial patterning in fixed cells that is underutilized in filamentous fungi. A primary complication for fixed-cell imaging in filamentous fungi is the need for enzymatic cell wall permeabilization, which is compounded by considerable variability in cell wall composition between species. smFISH adds another layer of complexity due to a requirement for RNase free conditions. Here, we describe the cloning, expression, and purification of a chitinase suitable for supplementation of a commercially available RNase-free enzyme preparation for efficient permeabilization of the Neurospora cell wall. We further provide a method for smFISH in Neurospora which includes a tool for generating numerical data from images that can be used in downstream customized analysis protocols.

Indexed as

Neurospora crassaCell WallDigestionRibonucleasesRNARibonucleasesRNAChitinaseFixed-cell imagingFungal microscopyQuantitative cell biology

Identifiers

PMID34425213
PMCPMC8463489
OpenAlexW3196166987

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.