CoChat
Sign Up
Cited by 12Congenital heart defects researchGenomics and Chromatin DynamicsDevelopmental Biology and Gene Regulation

The DOI matches its Crossref record; 75 of the 75 references it cites were found in a registry or an index; 37 of the 56 sources its claims cite were checked for support.

A cis-regulatory-directed pipeline for the identification of genes involved in cardiac development and disease

Open access
Nim, Hieu T., Dang, Louis, Thiyagarajah, Harshini, Bakopoulos, Daniel, See, Michael, Charitakis, Natalie, Sibbritt, Tennille, Eichenlaub, Michael P., Archer, Stuart K., Fossat, Nicolas, Burke, Richard E., Tam, Patrick P. L., Warr, Coral G., Johnson, Travis K., Ramialison, Mirana·2021·Genome Biology
DOI 10.1186/s13059-021-02539-0PMC PMC8672579
Open this paper in CoChat — free accountOpen PDF

Abstract

Background: Congenital heart diseases are the major cause of death in newborns, but the genetic etiology of this developmental disorder is not fully known. The conventional approach to identify the disease-causing genes focuses on screening genes that display heart-specific expression during development. However, this approach would have discounted genes that are expressed widely in other tissues but may play critical roles in heart development. Results: We report an efficient pipeline of genome-wide gene discovery based on the identification of a cardiac-specific cis-regulatory element signature that points to candidate genes involved in heart development and congenital heart disease. With this pipeline, we retrieve 76% of the known cardiac developmental genes and predict 35 novel genes that previously had no known connectivity to heart development. Functional validation of these novel cardiac genes by RNAi-mediated knockdown of the conserved orthologs in Drosophila cardiac tissue reveals that disrupting the activity of 71% of these genes leads to adult mortality. Among these genes, RpL14, RpS24, and Rpn8 are associated with heart phenotypes. Conclusions: Our pipeline has enabled the discovery of novel genes with roles in heart development. This workflow, which relies on screening for non-coding cis-regulatory signatures, is amenable for identifying developmental and disease genes for an organ without constraining to genes that are expressed exclusively in the organ of interest.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

The authors present a cis-regulatory-element-directed pipeline that predicts cardiac developmental and disease genes without requiring heart-specific gene expression, and validate novel candidates by cardiac RNAi knockdown in Drosophila. The pipeline recovers most known cardiac developmental genes and identifies 35 previously unlinked candidates, many of which affect adult viability or heart morphology when knocked down.

Key findings

  • The pipeline retrieved 76% of known cardiac developmental genes and predicted 35 novel genes with no prior known connectivity to heart development.
  • Of genes with both cardiac-specific promoters and enhancers (Category I), 26/44 (59%) known CHD-causing genes were recovered; overall 84% of known cardiac disease genes were associated with a cardiac-specific CRE.
  • In Drosophila cardiac RNAi knockdown, 71% of tested genes (17/24) were associated with reduced adult viability; RpL14, RpS24, and Rpn8 showed severe mortality and heart phenotypes including reduced pericardial cell size, heart tube defects, and loss of heartbeat.
  • Category I genes were enriched for heart development (top GO term GO:0007507), while Category IV genes with ubiquitous CREs were enriched for housekeeping functions such as translation.

Methods

  • Genome-wide bioinformatic pipeline using mouse E14.5 ENCODE ChIP-seq data for H3K4me3, H3K4me1, and H3K27ac across heart, limb, liver, and brain; cardiac-specific CREs were defined by heart-specific histone marks and assigned to genes with GREAT.
  • RNA-seq from mouse E14.5 heart was used to filter highly expressed genes (>20 FPKM, top 8% of 17,052 detected genes); network reconstruction used STRING v10 and Cytoscape.
  • Functional validation used Drosophila melanogaster RNAi knockdown driven by 4xHand-Gal4 from embryonic stage 13 onward, with mortality index, hatching rate, heart rate, confocal imaging, F-actin staining, and anti-Pericardin staining.

Limitations

  • The paper does not include a dedicated limitations section. It reports that the overlap between Category I genes and CHD de novo mutations was not statistically significant (11% vs 5% control, p = 0.057).
  • Three candidate genes (CG5885, CG8004, Oststt3) were not tested due to unavailable RNAi lines; the high-expression cutoff of 20 FPKM was described as arbitrary.
  • Evident from the text: the discovery pipeline uses only mouse E14.5 heart and three comparison tissues, and functional validation relies on Drosophila orthologs rather than human or mammalian cardiac models.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1186/s13059-021-02539-0

Cites

75 of 75 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

37 of 56 cited sources checked · 20 supported · 1 flagged · accuracy not yet measured

  • Findingown data

    Category IV showed enrichment of TFBSs for core transcription factors, such as Sp1.

    “In contrast, Category IV showed enrichment of TFBSs for core transcription factors, such as Sp1 which has essential roles in cell growth, differentiation, apoptosis, and carcinogenesis (Fig. 2C, iv) [27].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [27] Vizcaíno C, Mansilla S, Portugal J. Sp1 transcription factor: a long-standing target in cancer chemotherapy. Pharmacol Ther. 2015;152:111–24. https://doi.org/10.1016/j.pharmthera.2015.05.008. source
  • Findingown data

    A strong cardiac cis-regulatory signature was observed in Category I with enrichment of binding sites of known master regulators of cardiac development, such as Nkx2-5 and Tbx5.

    “A strong cardiac cis-regulatory signature was observed in Category I with the enrichment of binding sites of known master regulators of cardiac development, such as Nkx2-5 and Tbx5, which when mutated account for the genetic causes of CHD (Fig. 2C, i) [2, 25].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [2] Waardenberg AJ, Ramialison M, Bouveret R, Harvey RP. Genetic networks governing heart development. Cold Spring Harb Perspect Med. 2014;4(11). https://doi.org/10.1101/cshperspect.a013839. source

      The Cardiac Kernel section identifies NKX2-5 and TBX5 as important transcription factors in heart development, and the Enhancer Function section explains how transcription-factor binding sites contribute to cardiac enhancers. The source does not report a “Category I” group or enrichment of NKX2-5 and TBX5 binding sites in it; that observation is not supported by this paper. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [25] Bouveret R, Waardenberg AJ, Schonrock N, Ramialison M, Doan T, de Jong D, et al. NKX2-5 mutations causative for congenital heart disease retain functionality and are directed to hundreds of targets. Elife. 2015;4. https://doi.org/10.7554/eLife.06942. source

      The Results section on NKX2-5 mutant targets reports enrichment of NKX2-5 binding motifs in the paper’s A- and B-sets, and the Figure 1 caption identifies NKX2-5 mutations associated with congenital heart disease. The cited paper does not report the citing document’s Category I signature or enrichment of Tbx5 binding sites; its analyses concern different target sets. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
  • Interpretationown data

    The cCREs are possibly controlled by TFs of the cardiac kernel.

    “These results point to the possibility that the cCREs are controlled by TFs of the cardiac kernel [2].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [2] Waardenberg AJ, Ramialison M, Bouveret R, Harvey RP. Genetic networks governing heart development. Cold Spring Harb Perspect Med. 2014;4(11). https://doi.org/10.1101/cshperspect.a013839. source
  • Findingown data

    GO analysis revealed that the top enriched biological process term in the 1311 genes is heart development (GO:0007507).

    “Gene Ontology (GO) analysis using Metascape [22] revealed that the top enriched biological process term in this set is GO: 0007507: heart development”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [22] Zhou Y, Zhou B, Pache L, Chang M, Khodabakhshi AH, Tanaseichuk O, et al. Metascape provides a biologist-oriented resource for the analysis of systems-level datasets. Nat Commun. 2019;10(1):1523. https://doi.org/10.1038/s41467-019-09234-6. source
  • Premisecitation

    The function of most CREs is unknown.

    “The function of most CREs is unknown; however, there is mounting evidence that sequence mutations in cardiac CREs are associated with congenital heart disease [2, 3].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [2] Waardenberg AJ, Ramialison M, Bouveret R, Harvey RP. Genetic networks governing heart development. Cold Spring Harb Perspect Med. 2014;4(11). https://doi.org/10.1101/cshperspect.a013839. source

      The Enhancer Function section describes what enhancers do, while the genome-wide discussion says consortia are working to describe functional genomic elements. Neither says that the function of most cis-regulatory elements is unknown. I searched for that assertion and for equivalent statements about uncharacterized enhancers; the passage about functions only beginning to be understood refers to long noncoding RNAs, not CREs. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [3] Chahal G, Tyagi S, Ramialison M. Navigating the non-coding genome in heart development and congenital heart disease. Differentiation. 2019;107:11–23. https://doi.org/10.1016/j.diff.2019.05.001. source

      Section 2.6 (page unavailable) says putative cREs need functional validation and that key non-coding elements implicated in congenital heart disease remain unknown. This supports uncertainty about cardiac cREs in that context, but does not establish that the function of most cREs generally is unknown. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
  • Probably not a substantial claim. 16
  • It may state two separate claims as one. 37

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    The VISTA Cardiac Enhancer Browser contains 2870 cardiac enhancers.

    “This database contains 2870 cardiac enhancers that were also obtained by computational prediction, with some having been functionally validated.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [28] Spurrell CH, Barozzi I, Mannion BJ, Blow MJ, Fukuda-Yuzawa Y, Afzal SY, Akiyama JA, Afzal V, Tran S, Plajzer-Frick I, et al. Genome-Wide Fetalization of Enhancer Architecture in Heart Disease. 2019. Preprint at https://www.biorxiv.org/content/10.1101/591362v1. source

      In “Highly reproducible heart enhancer landscape in healthy individuals” (page unavailable), the paper says the VISTA Enhancer Browser contains 284 human and mouse loci with validated heart activity, not 2,870 cardiac enhancers. It describes those 284 as tested in transgenic mouse assays, rather than saying only some were functionally validated. (LLM, conf=0.90)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    A cardiac gene regulatory network regulates heart development from a simple tubular structure into a pump under electrophysiological control.

    “a cardiac gene regulatory network (GRN) regulates the development of the heart from a simple tubular structure into a pump under electrophysiological control [2].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [2] Waardenberg AJ, Ramialison M, Bouveret R, Harvey RP. Genetic networks governing heart development. Cold Spring Harb Perspect Med. 2014;4(11). https://doi.org/10.1101/cshperspect.a013839. source

  • Probably not a substantial claim. It may state two separate claims as one.

    Premisecitation

    Promoters located close to the TSS are associated with core transcriptional functions, while distally located CREs are associated with tissue-specific functions.

    “Promoters located close to the TSS are associated with core transcriptional functions [24], while distally located CREs are associated with tissue-specific functions [3].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [24] Haberle V, Stark A. Eukaryotic core promoters and the functional basis of transcription initiation. Nat Rev Mol Cell Biol. 2018;19(10):621–37. https://doi.org/10.1038/s41580-018-0028-8. source
    • cites [3] Chahal G, Tyagi S, Ramialison M. Navigating the non-coding genome in heart development and congenital heart disease. Differentiation. 2019;107:11–23. https://doi.org/10.1016/j.diff.2019.05.001. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    More than 100,000 putative cardiac CREs have been documented, characterized by H3K4me3, H3K4me1, and H3K27ac marks in embryonic and adult heart tissues of mouse and human.

    “which has documented more than 100,000 putative cardiac CREs that are characterized by H3K4me3, H3K4me1, and H3K27ac marks in embryonic, adult heart tissues of mouse and human (NIH Roadmap Epigenomics Mapping Consortium) [16, 17].”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [16] Bernstein BE, Stamatoyannopoulos JA, Costello JF, Ren B, Milosavljevic A, Meissner A, et al. The NIH Roadmap Epigenomics Mapping Consortium. Nat Biotechnol. 2010;28(10):1045–8. https://doi.org/10.1038/nbt1010-1045. source

      The 'Reference maps for major epigenomic features' section identifies H3K4me1 and H3K4me3 as marks selected for mapping, and the 'Prioritized cells and tissues' section discusses human fetal and adult tissues relevant to cardiovascular disease. The paper does not report documenting more than 100,000 cardiac CREs in mouse and human heart tissue, or characterize them using H3K27ac. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [17] Rivera CM, Ren B. Mapping human epigenomes. Cell. 2013;155(1):39–55. https://doi.org/10.1016/j.cell.2013.09.011. source

      The 'Annotation of cis-Regulatory Elements from Chromatin Profiles' section describes histone marks used to identify regulatory elements, and the 'Dynamic Chromatin Landscapes During Human Development' section mentions putative enhancers studied in cardiac development (pages unavailable in our copy). Neither section documents more than 100,000 cardiac CREs or specifies embryonic and adult heart tissues from both mice and humans. (LLM, conf=0.82)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    The origin of congenital heart disease is unknown in 80% of cases.

    “the origin of CHD is unknown in 80% of the cases, suggesting that several determinants of heart disease, including genetic, are yet to be identified [3].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [3] Chahal G, Tyagi S, Ramialison M. Navigating the non-coding genome in heart development and congenital heart disease. Differentiation. 2019;107:11–23. https://doi.org/10.1016/j.diff.2019.05.001. source

      The Introduction says the cause remained unknown for some familial CHD cohorts and that the majority of patients with familial CHD lack a genetic diagnosis. It does not say that the origin is unknown in 80% of all CHD cases; the cited figures concern familial cohorts, not all cases. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

  • It may state two separate claims as one.

    Premisecitation

    The Minute phenotype is characterized by developmental delay, impaired growth, poor fertility, and cardiac dysfunction.

    “The Minute phenotype is characterized by developmental delay, impaired growth, poor fertility, and also cardiac dysfunction [48]”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [48] Casad ME, Abraham D, Kim IM, Frangakis S, Dong B, Lin N, et al. Cardiomyopathy is associated with ribosomal protein gene haplo-insufficiency in Drosophila melanogaster. Genetics. 2011;189(3):861–70. https://doi.org/10.1534/genetics.111.131482. source

  • Probably not a substantial claim. It may state two separate claims as one.

    Premisecitation

    DBA is a dominant Minute-like condition characterized by growth impairment, bone marrow failure, and congenital malformations with a high penetrance of CHD (~30%).

    “a dominant Minute-like condition characterized by growth impairment, bone marrow failure, and congenital malformations with a high penetrance of CHD (~ 30%) [49, 50].”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [49] Vlachos A, Blanc L, Lipton JM. Diamond Blackfan anemia: a model for the translational approach to understanding human disease. Expert Rev Hematol. 2014;7(3):359–72. https://doi.org/10.1586/17474086.2014.897923. source
    • cites [50] Vlachos A, Osorio DS, Atsidaftos E, Kang J, Lababidi ML, Seiden HS, et al. Increased prevalence of congenital heart disease in children with diamond Blackfan anemia suggests unrecognized diamond Blackfan anemia as a cause of congenital heart disease in the general population: a report of the Diamond Blackfan Anemia Registry. Circ Genom Precis Med. 2018;11(5):e002044. https://doi.org/10.1161/CIRCGENETICS.117.002044. source

      The paper’s main text (page unavailable) describes DBA as a bone marrow failure syndrome with growth failure and birth defects, and says ribosomal-protein-associated DBA has autosomal dominant inheritance. It does not describe DBA as “Minute-like.” More importantly, its ~30% figure is the proportion of congenital anomalies that are CHD, not CHD penetrance among people with DBA; the registry reports CHD in 14.9% of DBA patients. (LLM, conf=0.93)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim.

    Premisecitation

    Systematic cardiac gene regulatory networks were previously reported based on spatio-temporal datasets or integration of gene expression and gene regulation.

    “Indeed, systematic cardiac GRNs were previously reported, based on either spatio-temporal datasets [44, 45] or integration of layers of gene expression and gene regulation [46].”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [44] Herrmann F, Groß A, Zhou D, Kestler HA, Kühl M. A boolean model of the cardiac gene regulatory network determining first and second heart field identity. PLoS One. 2012;7(10):e46798. https://doi.org/10.1371/journal.pone.0046798. source
    • cites [45] Bazil JN, Stamm KD, Li X, Thiagarajan R, Nelson TJ, Tomita-Mitchell A, et al. The inferred cardiogenic gene regulatory network in the mammalian heart. PLoS One. 2014;9(6):e100842. https://doi.org/10.1371/journal.pone.0100842. source

      The Introduction and Materials and Methods describe a systematic cardiogenic gene regulatory network inferred from developing-mouse-heart expression data. The method combines expression-based network inference with gene-ontology similarity, not with a separate layer of gene-regulation data, so the specific description of what was integrated is not supported. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [46] Gong W, Koyano-Nakagawa N, Li T, Garry DJ. Inferring dynamic gene regulatory networks in cardiac differentiation through the integration of multi-dimensional data. BMC Bioinformatics. 2015;16(1):74. https://doi.org/10.1186/s12859-015-0460-0. source

  • It may state two separate claims as one.

    Interpretationcitation

    The identified modules recapitulate known modules of the cardiac GRN: the heart development node corresponds to the known kernel of cardiac transcription factors, and other modules correspond to known downstream gene batteries.

    “These modules recapitulate known modules of the cardiac GRN: the “heart development” node corresponds to the known kernel of cardiac transcription factors [2]. Other modules such as “muscle gene battery,” “angiogenesis,” “cell cycle and cytoskeleton,” and “mitochondrial genes” modules correspond to known downstream gene batteries of the cardiac GRN [3].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [2] Waardenberg AJ, Ramialison M, Bouveret R, Harvey RP. Genetic networks governing heart development. Cold Spring Harb Perspect Med. 2014;4(11). https://doi.org/10.1101/cshperspect.a013839. source
    • cites [3] Chahal G, Tyagi S, Ramialison M. Navigating the non-coding genome in heart development and congenital heart disease. Differentiation. 2019;107:11–23. https://doi.org/10.1016/j.diff.2019.05.001. source

  • It may state two separate claims as one.

    Findingcitation

    Available information in the MGI database indicates that for some of these genes, their knock-out results in heart defects or they are expressed in the heart.

    “However, available information in the MGI database [32, 34, 36] indicates that for some of these genes, their knock-out results in heart defects (Fig. 4, red border) or they are expressed in the heart (Fig. 4, double-line border).”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [32] Smith CM, Finger JH, Hayamizu TF, McCright IJ, Xu J, Berghout J, et al. The mouse Gene Expression Database (GXD): 2014 update. Nucleic Acids Res. 2014;42(D1):D818–24. https://doi.org/10.1093/nar/gkt954. source

      The Introduction and the section on new search forms describe GXD as part of MGI and explain that users can search gene-expression data by anatomy and genes associated with phenotypes (page numbers unavailable). The paper does not identify the genes in the citing sentence or report that their knockouts cause heart defects or that they are expressed in the heart. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [34] Bult CJ, Blake JA, Smith CL, Kadin JA, Richardson JE. Mouse Genome Database (MGD) 2019. Nucleic Acids Res. 2019;47(D1):D801–d806. https://doi.org/10.1093/nar/gky1056. source

      The Introduction describes MGD as a source of genotype-to-phenotype annotations, and the Phenotype/Gene expression comparison matrix section says it displays tissues where genes are expressed and where mutations cause abnormal phenotypes. Neither passage identifies the citing paper’s genes or establishes that their knockouts cause heart defects or that they are expressed in the heart. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [36] Eppig JT, Blake JA, Bult CJ, Richardson JE, Kadin JA, Ringwald M. Mouse genome informatics (MGI) resources for pathology and toxicology. Toxicol Pathol. 2007;35(3):456–7. https://doi.org/10.1080/01926230701310536. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    RPL14 produces a dominant 'Minute-like' phenotype in a vertebrate model.

    “RPL14 is yet to be linked to human disease, but is also known to produce a dominant “Minute-like” phenotype in a vertebrate model [51].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [51] Lai K, Amsterdam A, Farrington S, Bronson RT, Hopkins N, Lees JA. Many ribosomal protein mutations are associated with growth impairment and tumor predisposition in zebrafish. Dev Dyn. 2009;238(1):76–85. https://doi.org/10.1002/dvdy.21815. source

  • Probably not a substantial claim.

    Interpretationcitation

    Given that ENCODE, Epigenetics Roadmap, and FANTOM consortia contain comprehensive datasets for a multitude of tissues and cell types at different developmental stages, it would be feasible to mine the datasets using this pipeline for other time points or tissues of interest.

    “Finally, while our pipeline was run with the goal to identify cardiac genes, given that the ENCODE [15], Epigenetics Roadmap [16], and FANTOM [52] consortia contain comprehensive datasets for a multitude of tissues and cell types at different developmental stages, it would be feasible to mine the datasets using this pipeline for other time points or tissues of interest.”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [15] ENCODE Project Consortium. An integrated encyclopedia of DNA elements in the human genome. Nature. 2012;489(7414):57–74. https://doi.org/10.1038/nature11247. source

      Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured
    • cites [16] Bernstein BE, Stamatoyannopoulos JA, Costello JF, Ren B, Milosavljevic A, Meissner A, et al. The NIH Roadmap Epigenomics Mapping Consortium. Nat Biotechnol. 2010;28(10):1045–8. https://doi.org/10.1038/nbt1010-1045. source
    • cites [52] Forrest AR, Kawaji H, Rehli M, Baillie JK, de Hoon MJ, Haberle V, et al. A promoter-level mammalian expression atlas. Nature. 2014;507(7493):462–70. https://doi.org/10.1038/nature13182. source

  • It may state two separate claims as one.

    Interpretationown data

    This hit rate is well above the reported hit rate (10%) of a genome-wide cardiac RNAi screen and comparable to the 53% of a targeted approach based on sequence information of CHD patients.

    “This is well above the reported hit rate (10%) of a genome-wide cardiac RNAi screen [43] and is comparable to the 53% of a targeted approach based on sequence information of CHD patients [42].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [43] Neely GG, Kuba K, Cammarato A, Isobe K, Amann S, Zhang L, et al. A global in vivo Drosophila RNAi screen identifies NOT3 as a conserved regulator of heart function. Cell. 2010;141(1):142–53. https://doi.org/10.1016/j.cell.2010.02.023. source

      The Results report that approximately 10% of 80 randomly selected RNAi lines in a prescreen began to die at increased temperature. The paper did conduct a genome-wide cardiac RNAi screen, but the cited 10% figure describes the prescreen, not a reported hit rate for the genome-wide screen. The 53% figure is attributed to a different citation and is not assessed here. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [42] Zhu J-y FY, Nettleton M, Richman A, Han Z. High throughput in vivo functional validation of candidate congenital heart disease genes in Drosophila. eLife. 2017;6:e22617. https://doi.org/10.7554/eLife.22617. source

  • It may state two separate claims as one.

    Findingown data

    Each gene was typically associated with 1 or 2 CREs and the number of associated CREs per gene was consistent across all categories.

    “Overall, each gene was typically associated with 1 or 2 CREs and the number of associated CREs per gene was consistent across all categories (Additional file 1: Fig S3).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    RpL14, RpS24, and Rpn8 are associated with heart phenotypes.

    “Among these genes, RpL14, RpS24, and Rpn8 are associated with heart phenotypes.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    cCREs of categories I, II, and III were mostly located further away from the TSS, >5 kb upstream for cardiac promoters, and >5 kb upstream and downstream for cardiac enhancers.

    “cCREs of categories I, II, and III were mostly located further away from the TSS, > 5 kb upstream for cardiac promoters (Fig. 2B, i,ii), and > 5 kb upstream and downstream for cardiac enhancers (Fig. 2B, i’,iii).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    The authors retrieved 137 human genes implicated in cardiac diseases, 44 of which have a mouse ortholog expressed at E14.5.

    “We retrieved 137 human genes implicated in cardiac diseases, 44 of which have a mouse ortholog expressed at E14.5.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingcitation

    Twenty-three of these orthologs were known to be expressed in fly cardiac tissue, while the expression of three of the remaining four has not been reported.

    “Twenty-three of these were known to be expressed in fly cardiac tissue [15], while the expression of three of the remaining four has not been reported (Fig. 5A).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [15] ENCODE Project Consortium. An integrated encyclopedia of DNA elements in the human genome. Nature. 2012;489(7414):57–74. https://doi.org/10.1038/nature11247. source

  • It may state two separate claims as one.

    Findingown data

    Out of the 153 matched genes, 92 were also detectably expressed in the brain, 108 in the limb, and 99 in the liver.

    “Out of these, 92 were also detectably expressed in the brain, 108 in the limb, and 99 in the liver (Fig. 3A).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Staining for F-Actin and anti-Pericardin revealed defects including partially open heart tubes (RpL14 and Rpn8), and heart atrophy (RpS24).

    “Staining for F-Actin (to label heart tube filaments) and anti-Pericardin (cardiac collagen) revealed defects including partially open heart tubes (RpL14 and Rpn8), and heart atrophy (RpS24, Fig. 6D).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Genes associated with either one cardiac-specific promoter (Category II) or one enhancer (Category III) retained enrichment with muscle function while also displaying enrichment in generic functions.

    “Genes associated with either one cardiac-specific promoter (Category II) or one enhancer (Category III) retained an enrichment with muscle function (e.g., GO:0003012: muscle system process), while also displaying an enrichment in generic functions (e.g., GO:0006091: generation of precursor metabolites and energy, which is also shared with Category IV) (Fig. 2D).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    RpL14 and RpS24 knockdown larvae have diminished pericardial cell size, and Rpn8 knockdown larvae have cell size variability.

    “Note the diminished pericardial cell size in RpL14 and RpS24 knockdown larvae and cell size variability in Rpn8 knockdown larvae”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Cardiac knockdown of RpL14, RpS24, or Rpn8 in third instar larvae displayed partial or complete heart tube atrophy and infiltration of cells likely macrophages.

    “Knockdowns displayed partial (RpL14, Rpn8, arrowed) or complete heart tube atrophy (RpS24) and infiltration of cells (likely macrophages, asterisks).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Cardiac RNAi against RpL14, RpS24, and Rpn8 does not affect embryo hatching rates nor first instar heart rates relative to sibling controls.

    “Cardiac RNAi against RpL14, RpS24, and Rpn8 does not affect embryo hatching rates (A) nor first instar heart rates (B) relative to sibling controls. ns, not significant.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Newly hatched larvae showed a normal organization and size of pericardial cells of the heart tube and an unaffected heartbeat.

    “Newly hatched larvae showed a normal organization and size of pericardial cells of the heart tube and an unaffected heartbeat (Fig. 6B; RpL14: p = 0.659; RpS24: p = 0.058; Rpn8: p = 0.919).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Cardiac collagen was thickened and often broken in RpS24 knockdown, thinner and sparser in RpL14 knockdown, and relatively normal in Rpn8 knockdown.

    “cardiac collagen was thickened and often broken (RpS24), thinner and sparser (RpL14), and relatively normal (Rpn8).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    RNAi-expressing embryos hatched into larvae at similar proportions to their controls, indicating that mortality occurred during the post-embryonic stages.

    “RNAi-expressing embryos hatched into larvae at similar proportions to their controls, indicating that mortality occurred during the post-embryonic stages (Fig. 6A).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Findingown data

    A total of 153 genes could be mapped across the four tissue samples (limb, liver, brain, heart).

    “A total of 153 genes could be mapped across the 4 tissue samples.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    RNAi-knockdown of RpS24, RpL14, and Rpn8 led to complete or partly penetrant adult mortality and variably reduced viability for 14 other orthologs.

    “RNAi-knockdown of RpS24, RpL14, and Rpn8 (ortholog for the mouse Psmd7 gene) led to complete or partly penetrant adult mortality and variably reduced viability for 14 other orthologs (Fig. 5).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Ppp1r3c shows regionalized cardiac expression by in situ hybridization in murine heart section at E14.5, but also displays weaker expression in other tissues.

    “Ppp1r3c, for instance, shows regionalized cardiac expression by in situ hybridization in murine heart section at E14.5 (Fig. 3E), but it also displays weaker expression in other tissues (not observed in the cross-tissue RNA-seq data (Fig. 3A), likely owing to the presence of both a uCRE and a cCRE at its promoter region (Fig. 3E).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Closer inspection revealed strongly reduced pericardial cell size, with Rpn8 knockdown larvae also showing inconsistently sized cells.

    “Closer inspection revealed strongly reduced pericardial cell size, with Rpn8 knockdown larvae also showing inconsistently sized cells (Fig. 6C).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    48 h later at the third instar stage, the hearts of these larvae had ceased contracting and the cardiac cells were often absent.

    “However, 48 h later at the third instar stage, the hearts of these larvae had ceased contracting and the cardiac cells were often absent.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingcitation

    Cbx5 is highly expressed in all four tissues and is widely expressed in E14.5 embryo.

    “Similarly, Cbx5 is also highly expressed in all four tissues (Fig. 3A) and is widely expressed in E14.5 embryo [34] (Fig. 3D).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [34] Bult CJ, Blake JA, Smith CL, Kadin JA, Richardson JE. Mouse Genome Database (MGD) 2019. Nucleic Acids Res. 2019;47(D1):D801–d806. https://doi.org/10.1093/nar/gky1056. source

  • It may state two separate claims as one.

    Findingown data

    Category I genes were most enriched in cardiac and muscle development categories, with the top enriched term being GO:0007507 heart development.

    “We found that Category I genes were most enriched in cardiac and muscle development categories (e.g., the top enriched term was GO:0007507: heart development) (Fig. 2D)”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingcitation

    The majority of these exclusively heart-expressed genes (22/26) have been shown to be involved in heart development and disease and include known cardiac GRN components such as Nkx2-5, Myh6, Myl2, Myl4, and Nppb.

    “The majority of these genes (22/26) have been shown to be involved in heart development and disease and includes well-known cardiac GRN components such as Nkx2-5 (Fig. 3A), Myh6, Myl2, Myl4, and Nppb [2].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [2] Waardenberg AJ, Ramialison M, Bouveret R, Harvey RP. Genetic networks governing heart development. Cold Spring Harb Perspect Med. 2014;4(11). https://doi.org/10.1101/cshperspect.a013839. source

      Checked before — its earlier verdict is held back until a new check runs.

  • It may state two separate claims as one.

    Findingown data

    In RpS24 knockdown, Pericardin is thickened with visible breaks; in RpL14 knockdown, collagen appears thin and less dense.

    “Pericardin is thickened and breaks are visible for RpS24 (arrowhead), while for RpL14 collagen appears thin and less dense.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Findingcitation

    In total, 17,052 genes had detectable expression (FPKM_avg > 0) in the mouse embryonic heart at E14.5.

    “In total, 17,052 genes had detectable expression (FPKM_avg > 0) [30].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [30] GitHub: cardiac network component predictor using cis-regulatory elements (CREs) [https://github.com/Ramialison-Lab/CardiacNetworkComponentPredictor]. Accessed 2021.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • It may state two separate claims as one.

    Findingown data

    Third instar larvae with cardiac knockdown of RpL14, RpS24, or Rpn8 had no heartbeat.

    “These larvae had no heartbeat.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Interpretationreasoning

    The pipeline is readily applicable to other organs to identify novel GRN components.

    “This pipeline is readily applicable to other organs to identify novel GRN components.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Interpretationown data

    Predicted cardiac cCREs, in particular those associated with Category I genes, are able to deliver cardiac-specific functions, in contrast to uCREs that would drive non-tissue-specific functions.

    “Altogether, these results support that our predicted cardiac cCREs, in particular those associated with Category I genes, are able to deliver cardiac-specific functions, in contrast to uCREs that would drive non-tissue-specific functions.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationown data

    The pipeline complements the conventional approach and allows the identification of novel heart development genes that are not expressed specifically in the heart.

    “Together, our results support that this pipeline complements the conventional approach and allows the identification of novel heart development genes that are not expressed specifically in the heart.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationown data

    RpL14, RpS24, and Rpn8 are essential for pericardial cell function and cardiac integrity.

    “These phenotypes suggest that RpL14, RpS24, and Rpn8 are essential for pericardial cell function and cardiac integrity.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationreasoning

    The fly heart is disproportionately sensitive to reduced ribosomal function, likely explaining the severe phenotypes observed upon RNAi knockdown.

    “suggesting that the fly heart is disproportionately sensitive to reduced ribosomal function; likely why we observed such severe phenotypes upon RNAi knockdown.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Atp2a2 is involved in the regulation of cardiac muscle contraction and is regulated by the cardiac TF TBX5.

    “For example, Atp2a2 is a gene involved in the regulation of cardiac muscle contraction and is regulated by the cardiac TF TBX5 [33].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [33] Zhu Y, Gramolini AO, Walsh MA, Zhou YQ, Slorach C, Friedberg MK, et al. Tbx5-dependent pathway regulating diastolic function in congenital heart disease. Proc Natl Acad Sci U S A. 2008;105(14):5519–24. https://doi.org/10.1073/pnas.0801779105. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Mutations in human ribosomal proteins, including RPS24, cause Diamond-Blackfan anemia (DBA).

    “mutations in human ribosomal proteins (including RPS24) cause Diamond-Blackfan anemia (DBA, RPS24 is DBA3 MIM:610629)”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [49] Vlachos A, Blanc L, Lipton JM. Diamond Blackfan anemia: a model for the translational approach to understanding human disease. Expert Rev Hematol. 2014;7(3):359–72. https://doi.org/10.1586/17474086.2014.897923. source
    • cites [50] Vlachos A, Osorio DS, Atsidaftos E, Kang J, Lababidi ML, Seiden HS, et al. Increased prevalence of congenital heart disease in children with diamond Blackfan anemia suggests unrecognized diamond Blackfan anemia as a cause of congenital heart disease in the general population: a report of the Diamond Blackfan Anemia Registry. Circ Genom Precis Med. 2018;11(5):e002044. https://doi.org/10.1161/CIRCGENETICS.117.002044. source

      Checked before — its earlier verdict is held back until a new check runs.

  • Probably not a substantial claim.

    Interpretationnone given

    The approach capitalizes on the ever-growing wealth of gene regulatory information available in public repositories.

    “This approach capitalizes on the ever-growing wealth of gene regulatory information available in public repositories.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationown data

    Our approach is effective and highly complementary to existing approaches that rely on gene expression.

    “We present evidence demonstrating that our approach is both effective and highly complementary to existing approaches that rely on gene expression.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductown data

    Three genes (CG5885, CG8004, and Oststt3) were not tested due to unavailability of RNAi lines.

    “Three genes (CG5885, CG8004, and Oststt3) were not tested due to unavailability of RNAi lines.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Interpretationown data

    Both Atp2a2 and Cbx5 harbor a cCRE adjacent to the ubiquitous H3K4me3 mark, suggesting they are subjected to cardiac-specific regulation.

    “However, both genes also harbor a cCRE adjacent to the ubiquitous H3K4me3 mark (Fig. 3C, D), suggesting that they are subjected to cardiac-specific regulation.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationown data

    The novel genes identified by the pipeline do not display a cardiac-restricted/specific expression profile and would be overlooked in gene discovery pipelines based on heart-specific gene expression.

    “As these genes do not display a cardiac-restricted/specific expression profile, they would be overlooked in gene discovery pipelines based on knowledge of heart-specific gene expression.”
    Quote in the paperSubstantial claim
50 of 50

  • The paper relies on this without support.

    Conventional gene discovery for heart development relies on expression pattern profiling and validation by forward or reverse genetic approaches.

    “Identifying genes for heart development conventionally relies on expression pattern profiling, and validation by forward or reverse genetic approaches.”

  • The paper relies on this without support.

    Rpn8/PSMD7 encodes a key regulatory component of the 26S ubiquitin proteosome complex and is not linked to human disease.

    “Rpn8/PSMD7, which encodes a key regulatory component of the 26S ubiquitin proteosome complex, is also not linked to human disease.”

  • The paper relies on this without support.

    De novo mutations in Rpn5/PSMD12 cause Stankiewicz-Isidor syndrome, a neurodevelopmental disorder with a strong congenital cardiac malformation component.

    “Notably however, de novo mutations in an interacting subunit, Rpn5/PSMD12, causes Stankiewicz-Isidor syndrome; a recently discovered neurodevelopmental disorder with a strong congenital cardiac malformation component.”

  • The paper relies on this without support.

    RPL14 has not been linked to human disease.

    “RPL14 is yet to be linked to human disease, but is also known to produce a dominant “Minute-like” phenotype in a vertebrate model [51].”
4 of 4

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Methods identify a different histone mark for enhancers

    Minor

    The Results say enhancers were identified by overlapping H3K4me1 and H3K27ac marks, but the Methods specify H3K4me3 and H3K27ac. These describe different inputs for the same enhancer-identification step.

1 of 1

Citations

Cites

Flagged first, then by citations

Research works printed but not linked: 7

  1. ENCODE Project Consortium. Total RNA-seq from heart (ENCSR727FHP): GSE78441. Gene Expression Omnibus; 2021. [cited 2021 Nov 1]. Available from: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE78441
  2. ENCODE Project Consortium: LICR_RnaSeq_Heart_E14.5. GSM929724. Gene Expression Omnibus; 2021 [cited 2021 Nov 1]. Available from: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM929724
  3. ENCODE Project Consortium: LICR_RnaSeq_Limb_E14.5. GSM929713. Gene Expression Omnibus; 2021 [cited 2021 Nov 1]. Available from: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM929713
  4. ENCODE Project Consortium: LICR_RnaSeq_Liver_E14.5. GSM929721. Gene Expression Omnibus; 2021 [cited 2021 Nov 1]. Available from: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM929721
  5. ENCODE Project Consortium: LICR_RnaSeq_WholeBrain_E14.5. GSM929723. Gene Expression Omnibus; 2021 [cited 2021 Nov 1]. Available from: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM929723
  6. Smith CM, Hayamizu TF, Finger JH, Bello SM, McCright IJ, Xu J, Baldarelli RM, Beal JS, Campbell JW, Corbani LE, Frost PJ, Lewis, JR, Giannatto SC, Miers DB, Shaw DR, Kadin JA, Richardson JE, Smith CL, Ringwald M. The mouse Gene Expression Database (GXD): 2019 update. MGI:5331042. MGI Gene Expression Database; 2021 [cited 2021 Nov 1]. Available from: http://www.informatics.jax.org/image/MGI:5331042
  7. Smith CM, Hayamizu TF, Finger JH, Bello SM, McCright IJ, Xu J, Baldarelli RM, Beal JS, Campbell JW, Corbani LE, Frost PJ, Lewis, JR, Giannatto SC, Miers DB, Shaw DR, Kadin JA, Richardson JE, Smith CL, Ringwald M. The mouse Gene Expression Database (GXD): 2019 update. MGI:4468106. MGI Gene Expression Database; 2021 [cited 2021 Nov 1]. Available from: http://www.informatics.jax.org/image/MGI:4468106

Cited by

Sorted by citations

Related

Metadata

Authors
Nim, H. T., Dang, L., Thiyagarajah, H., Bakopoulos, D., See, M., Charitakis, N., Sibbritt, T., Eichenlaub, M. P., Archer, S. K., Fossat, N., Burke, R. E., Tam, P. P. L., Warr, C. G., Johnson, T. K., & Ramialison, M.
Affiliations
  • Australian Regenerative Medicine Institute (AU)
  • Monash University (AU)
  • Murdoch Children's Research Institute (AU)
  • The University of Melbourne (AU)
  • The University of Sydney (AU)
  • University of Copenhagen (DK)
  • La Trobe University (AU)
Corresponding author
  • Nim, Hieu T.
  • Dang, Louis
  • Thiyagarajah, Harshini
  • Bakopoulos, Daniel
  • See, Michael
  • Charitakis, Natalie
  • Sibbritt, Tennille
  • Eichenlaub, Michael P.
  • Archer, Stuart K.
  • Fossat, Nicolas
  • Burke, Richard E.
  • Tam, Patrick P. L.
  • Warr, Coral G.
  • Johnson, Travis K.
  • Ramialison, Mirana
ORCID
Venue
Genome Biology
Publisher
Springer Science and Business Media LLC
Published
2021-12
Volume
22
Issue
1
Pages
335-335
ISSN
  • 1465-6906 (ISSN-L)
  • 1474-760X
DOI
10.1186/s13059-021-02539-0
PMC
PMC8672579
OpenAlex
W4200244660
Semantic Scholar
91eabbb50e371bdb2d8a3e7eae5b25ee7e82c26d
Cited by
11
References
74
Citations by year
2025: 6 · 2024: 1 · 2023: 4
FWCI
0.42
Type
journal article
Language
en
Open access
Licence
Locations
  • Genome biology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Minerva Access (University of Melbourne) (submittedVersion, cc-by)
Funders
  • Australian Research Council (DP140101067, DP160100933)
  • National Health and Medical Research Council (1049980, 1182330, 1180905)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Congenital heart defects research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
Keywords
heart development, congenital heart disease, cis-regulatory elements, candidate genes, Drosophila heart, RNAi knockdown, RPL14, RPS24, ribosomal protein genes
MeSH terms
Animals, Drosophila, Genetic Testing, Heart, Heart Defects, Congenital, Ribosomal Proteins, Genome, Gene Expression Regulation, Developmental, Computational Biology, Genomics, RNA Interference, Regulatory Elements, Transcriptional

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

A language model drew the claims from the paper on 2026-11-09, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-11-09 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Cited sources are checked with a language model’s help (gpt-6-sol).

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review