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Record W4312213379 · doi:10.1101/2022.12.27.522013

Improved genetically encoded fluorescent biosensors for monitoring of intra- and extracellular L-lactate

2022· preprint· en· W4312213379 on OpenAlexaff
Yusuke Nasu, Abhi Aggarwal, Giang N. T. Le, Yuki Kamijo, Marc Boisvert, Marie-Ève Paquet, Mikhail Drobizhev, Kaspar Podgorski, Robert E. Campbell

Bibliographic record

VenuebioRxiv (Cold Spring Harbor Laboratory) · 2022
Typepreprint
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicReceptor Mechanisms and Signaling
Canadian institutionsUniversity of AlbertaUniversité LavalUniversity of Toronto
FundersPrecursory Research for Embryonic Science and TechnologyJapan Society for the Promotion of ScienceNational Institutes of HealthToyota Physical and Chemical Research InstituteUniversity of TokyoJapan Science and Technology AgencyPrecise Measurement Technology Promotion FoundationMontana State University
KeywordsExtracellularFluorescenceIntracellularBiosensorBiochemistryGreen fluorescent proteinMetaboliteBiophysicsIn vitroMembraneBiologyChemistryCell biologyGene

Abstract

fetched live from OpenAlex

Abstract l -Lactate is increasingly appreciated as a key metabolite and signaling molecule in mammals. To enable investigations of both the inter- and intra-cellular dynamics of l -Lactate, we develop a second-generation green fluorescent extracellular l -Lactate biosensor, designated eLACCO2.1, and a red fluorescent intracellular l -Lactate biosensor, designated R-iLACCO1. Compared to the first-generation eLACCO1.1 (Δ F / F = 1.5 in cultured neurons), eLACCO2.1 exhibits better membrane localization and fluorescence response (Δ F / F = 8.1 in cultured neurons) with faster response kinetics to extracellular l -Lactate on the surface of live mammalian cells. R-iLACCO1 and its affinity variants exhibit large fluorescence responses to changes in l -Lactate concentration in vitro (Δ F / F = 15 to 22) and in live mammalian cells (Δ F / F = 5.5 to 11). We demonstrate that these biosensors enable cellular-resolution imaging of extracellular and intracellular l -Lactate in cultured mammalian cells.

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)0.001
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: Empirical · Consensus signal: Empirical
Teacher disagreement score0.001
Threshold uncertainty score0.005

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.001
Meta-epidemiology (narrow)0.0010.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0010.000
Research integrity0.0010.001
Insufficient payload (model declined to judge)0.0000.000

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 head0.011
GPT teacher head0.223
Teacher spread0.212 · 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
GenreEmpirical

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

Citations2
Published2022
Admission routes1
Has abstractyes

Explore more

Same venuebioRxiv (Cold Spring Harbor Laboratory)→Same topicReceptor Mechanisms and Signaling→French-language works237,207→