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Record W2418505177 · doi:10.1007/978-1-61779-005-8_13

Visualizing mRNAs in Fixed and Living Yeast Cells

2011· article· en· W2418505177 on OpenAlexafffund
Franck Gallardo, Pascal Chartrand

Bibliographic record

VenueMethods in molecular biology · 2011
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicRNA Research and Splicing
Canadian institutionsUniversité de Montréal
FundersCanadian Institutes of Health Research
KeywordsMessenger RNAGreen fluorescent proteinCell biologyIn situ hybridizationBiologyTranslation (biology)Saccharomyces cerevisiaeRNAP-bodiesYeastMolecular biologyBiochemistryGene

Abstract

fetched live from OpenAlex

Localization of messenger RNA (mRNA) is a process used by eukaryotes to control the spatio-temporal expression of proteins involved in cellular motility, asymmetric cell division, or polarized cell growth. A better understanding of this process relies on methods to detect specifically the position of an mRNA in fixed or living cells. This chapter presents methods to visualize mRNA in both fixed and living yeast Saccharomyces cerevisiae. In fixed cells, position of mRNAs can be assessed by using Fluorescent In Situ Hybridization (FISH) that consists of the hybridization of fluorescent probes that target a specific transcript in situ. In living cells, dynamics of mRNAs can be monitored using a bipartite system composed of MS2 stem-loops inserted in the mRNA of interest. These stem-loops are recognized specifically by the MS2 RNA-binding protein, fused to a fluorescent protein. In vivo association between the reporter (fluorescent MS2 protein) and the MS2-tagged mRNA reconstitutes active fluorescent ribonucleoparticles that can be followed by live cell imaging. Detailed protocols for the realization of these methods are provided and several technical considerations are discussed. Together, these methods provide very robust tools to determine the intracellular position and dynamics of your mRNA of interest in yeast.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Methods · Consensus signal: none
Teacher disagreement score0.003
Threshold uncertainty score0.010

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0010.000
Open science0.0010.000
Research integrity0.0000.001
Insufficient payload (model declined to judge)0.0030.002

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.037
GPT teacher head0.400
Teacher spread0.362 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreMethods

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations8
Published2011
Admission routes2
Has abstractyes

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