PreprintCited by 4Genomics and Chromatin DynamicsGene Regulatory Network AnalysisFungal and yeast genetics research

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

Predicting gene expression using millions of yeast promoters reveals cis -regulatory logic

Open access
Dash, Tirtharaj, Bornelöv, Susanne·2025·Bioinformatics Advances

Preprint — published as 10.1093/bioadv/vbaf130 · Paper page

Abstract

Motivation: Gene regulation involves complex interactions between multiple transcription factors. While early attempts to train deep neural networks to predict gene expression were limited to naturally occurring promoter sequences, the advent of gigantic parallel reporter assays has expanded available training data by orders of magnitude. Despite these advances, a clear understanding of how to use deep learning to study gene regulation is still lacking. Method: Here we investigate the complex association between gene promoters and expression in S. cerevisiae using Camformer, a residual convolutional neural network that ranked 4th in the Random Promoter DREAM Challenge 2022. We present the original model trained on 6.7 million random promoter sequences and investigate 270 alternative models to determine what factors contribute most to model performance. Finally, we use explainable AI to uncover regulatory signals. Results: We show that Camformer accurately decodes the association between promoters and gene expression ( r 2 = 0.914 ± 0.003, ρ = 0.962 ± 0.002) and provides a substantial improvement over previous state of the art. Furthermore, we show that a much smaller model with approximately 90% fewer parameters than the original model can achieve a high predictive performance. Using Grad-CAM and in silico mutagenesis, we demonstrate that the model learns both individual motifs and their hierarchy. For example, while an IME1 motif on its own increases gene expression, the co-occurrence of a UME6 motif provides a switch to strongly reduce gene expression. Thus, deep learning models such as Camformer can provide detailed insights into cis -regulatory logic. Availability and Implementation: The data and code used and developed in our experiments are publicly available at: https://github.com/Bornelov-lab/Camformer .

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1101/2025.01.25.634853

Cites

1 of 1 references verified0 citation errors · 0 not checkable

Statistics

Figures

Figure 1. Schematic of prediction task: (A) training data are arranged as (promoter, expression) pairs, (B) histogram of gene expression in training set, (C) the task is to predict YFP expression given a promoter sequence, and (D) promoter sequences are composed of an 80 nt random sequence positioned between two fixed sequences.
Figure 1. Schematic of prediction task: (A) training data are arranged as (promoter, expression) pairs, (B) histogram of gene expression in training set, (C) the task is to predict YFP expression given a promoter sequence, and (D) promoter sequences are composed of an 80 nt random sequence positioned between two fixed sequences.CC BY · PubMed Central
Figure 2. The final model consists of six convolutional layers followed by fully connected layers. Inputs and activations are shown in the format (h,w,c).
Figure 2. The final model consists of six convolutional layers followed by fully connected layers. Inputs and activations are shown in the format (h,w,c).CC BY · PubMed Central
Figure 3. The model was evaluated on the DREAM2022 test data (n = 71 103). (A) Scatter plot between true and predicted expression values. (B) Predictive performance measured using Pearson r² across 10 replicate models and ensemble models constructed by taking the mean prediction from k replicates (where k∈{2,…,10}). A replicate here refers to a model constructed using a random seed. (C) Same as (B), but showing Spearman ρ.
Figure 3. The model was evaluated on the DREAM2022 test data (n = 71 103). (A) Scatter plot between true and predicted expression values. (B) Predictive performance measured using Pearson r² across 10 replicate models and ensemble models constructed by taking the mean prediction from k replicates (where k∈{2,…,10}). A replicate here refers to a model constructed using a random seed. (C) Same as (B), but showing Spearman ρ.CC BY · PubMed Central
Figure 4. Evaluation and benchmark. (A, B) Camformer (Cam) and LegNet (Leg) performance across (A) challenging sequences, which displayed the strongest improvement over baseline (Vaishnav_deBoer; dashed grey line), or (B) SNVs, which displayed the weakest improvement over baseline. The top five DREAM2022 submissions are shown as coloured dashed lines. All sequence categories are shown in Figure S5. (C, D) Performance of Camformer in predicting expression for yeast grown in (C) complex or (D) defined medium. Performance of Camformer-Mini and LegNet is indicated. (E, F) Comparison of Camformer and Camformer-Mini with SOTA models. Prediction of expression for (E) native or (F) random promoters for yeast grown in complex medium. Violin plots show bootstraps with n = 10 000 replicates. All differences were significant (P<.05, paired Student’s t-test).
Figure 4. Evaluation and benchmark. (A, B) Camformer (Cam) and LegNet (Leg) performance across (A) challenging sequences, which displayed the strongest improvement over baseline (Vaishnav_deBoer; dashed grey line), or (B) SNVs, which displayed the weakest improvement over baseline. The top five DREAM2022 submissions are shown as coloured dashed lines. All sequence categories are shown in Figure S5. (C, D) Performance of Camformer in predicting expression for yeast grown in (C) complex or (D) defined medium. Performance of Camformer-Mini and LegNet is indicated. (E, F) Comparison of Camformer and Camformer-Mini with SOTA models. Prediction of expression for (E) native or (F) random promoters for yeast grown in complex medium. Violin plots show bootstraps with n = 10 000 replicates. All differences were significant (P<.05, paired Student’s t-test).CC BY · PubMed Central
Figure 5. t-SNE embedding for 10 000 promoters from the test set. The scatter plots show the t-SNE embeddings of the activations in the last convolutional layer. (A) The points are coloured based on their expression quantiles, where Q1 is the lowest and Q6 is the highest. (B) The points are coloured based on their error quantiles from very low to very high error. Embeddings for all layers are shown in Fig. S9.
Figure 5. t-SNE embedding for 10 000 promoters from the test set. The scatter plots show the t-SNE embeddings of the activations in the last convolutional layer. (A) The points are coloured based on their expression quantiles, where Q1 is the lowest and Q6 is the highest. (B) The points are coloured based on their error quantiles from very low to very high error. Embeddings for all layers are shown in Fig. S9.CC BY · PubMed Central
Figure 6. Camformer predictions reveal gene-regulatory logic. (A) The training data were divided into 10 expression quantiles (Q1 to Q10) and SEA was used to identify the most enriched motifs among the highest (Q10) and lowest (Q1) expressed promoters. (B) Heatmap showing mean predicted expression for training data sequences containing pairwise combinations of selected motifs as detected by FIMO. Baseline expressions for each individual motif are shown along the diagonal. (C) Heatmap showing mean expression difference between sequences with one motif (indicated by row) and sequences with two co-occurring motifs (row and column). (D) Heatmap showing correlation between predicted expression and motif count, score, or position. Grey values indicate cases where all motifs had the same score. (E) JASPAR2024 motif logos show that IME1 and UME6 motifs may overlap. (F) Venn diagram illustrating that some test set sequences had both motifs. (G) Heatmap showing ISM scores per position across IME1 motifs in the test set. The IME1 motifs are grouped into those that co-occur with UME6 (n=365) and those that appear on their own (n=2177). (H) Examples of motifs highlighted using ISM across variable positions 18–97. Positive values indicate contribution to higher expression and negative values indicate a contribution to lower expression. Positions and orientation of motifs are shown as arrows.
Figure 6. Camformer predictions reveal gene-regulatory logic. (A) The training data were divided into 10 expression quantiles (Q1 to Q10) and SEA was used to identify the most enriched motifs among the highest (Q10) and lowest (Q1) expressed promoters. (B) Heatmap showing mean predicted expression for training data sequences containing pairwise combinations of selected motifs as detected by FIMO. Baseline expressions for each individual motif are shown along the diagonal. (C) Heatmap showing mean expression difference between sequences with one motif (indicated by row) and sequences with two co-occurring motifs (row and column). (D) Heatmap showing correlation between predicted expression and motif count, score, or position. Grey values indicate cases where all motifs had the same score. (E) JASPAR2024 motif logos show that IME1 and UME6 motifs may overlap. (F) Venn diagram illustrating that some test set sequences had both motifs. (G) Heatmap showing ISM scores per position across IME1 motifs in the test set. The IME1 motifs are grouped into those that co-occur with UME6 (n=365) and those that appear on their own (n=2177). (H) Examples of motifs highlighted using ISM across variable positions 18–97. Positive values indicate contribution to higher expression and negative values indicate a contribution to lower expression. Positions and orientation of motifs are shown as arrows.CC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Self-check

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

The self-check needs the paper’s full text.

Citations

Cites

Show all 1 references

Research works printed but not linked: 27

  1. Takuya Akiba , Shotaro Sano , Toshihiko Yanase , et al. Optuna: A next-generation hyperparameter optimization framework. In Proc 25th ACM SIGKDD Int Conf Knowl Discov Data Min, pages 2623–2631, 2019. · DOI
  2. 10.1038/s41592-021-01252-x · DOI · Paper page
  3. 10.1038/S41588-021-00782-6 · DOI · Paper page
  4. 10.1038/s41587-019-0315-8 · DOI · Paper page
  5. 10.3389/fdata.2023.1113402 · DOI
  6. 10.1038/nature11245 · DOI
  7. 10.1371/journal.pcbi.1002592 · DOI
  8. 10.1093/genetics/iyad123 · DOI
  9. 10.1016/j.cell.2009.02.043 · DOI
  10. 10.1016/j.patrec.2021.06.030 · DOI
  11. 10.1186/s12859-024-05645-5 · DOI
  12. 2020, DeepATT: a hybrid category attention neural network for identifying functional effects of DNA sequences, Brief Bioinform, 22, bbaa159
  13. 10.1016/j.aiopen.2022.10.001 · DOI · Paper page
  14. 10.1016/j.cels.2021.05.015 · DOI
  15. 10.1038/nbt.1589 · DOI · Paper page
  16. 10.1093/bioinformatics/btad457 · DOI
  17. 10.1093/nar/gkw226 · DOI
  18. Abdul Muntakim Rafi , Daria Nogina , Dmitry Penzar , et al. A community effort to optimize sequence-based deep learning models of gene regulation. Nat Biotechnol, pages 1–11, 2024.
  19. Ramprasaath R Selvaraju , Michael Cogswell , Abhishek Das , et al. Grad-CAM: Visual explanations from deep networks via gradient-based localization. In Proc IEEE Int Conf Comput Vis, pages 618–626, 2017. · DOI · Paper page
  20. 2021, Efficientnetv2: Smaller models and faster training, arXiv preprint
  21. 10.1038/s41586-022-04506-6 · DOI
  22. Ashish Vaswani , Noam Shazeer , Niki Parmar , et al. Attention is all you need. Adv Neural Inf Process Syst, 30, 2017.
  23. Olivia Wiles , Sven Gowal , Florian Stimberg , et al. A fine-grained analysis on distribution shift. In International Conference on Learning Representations, 2022.
  24. 10.1038/s42256-021-00316-z · DOI
  25. 10.1038/nmeth.3547 · DOI · Paper page
  26. 2023, Xai meets biology: A comprehensive review of explainable ai in bioinformatics applications, arXiv preprint
  27. 10.1038/s41467-020-19921-4 · DOI · Paper page

Metadata

Authors
Dash, T., & Bornelöv, S.
Affiliations
University of Cambridge (GB)
Corresponding author
Bornelöv, Susanne
Venue
Bioinformatics Advances
Publisher
openRxiv
Published
2025-01-27
Volume
5
Issue
1
Pages
vbaf130-vbaf130
ISSN
2635-0041 (ISSN-L)
OpenAlex
W4410942367
Cited by
4
References
1
Citations by year
2026: 3 · 2025: 1
FWCI
0.54
Type
journal article
Language
en
Open access
Locations
  • Bioinformatics Advances (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Apollo (University of Cambridge) (submittedVersion, cc-by)
  • Apollo (cc-by)
Funders
  • Wellcome Trust (226518/Z/22/Z, 226518)
  • Dell EMC
  • Engineering and Physical Sciences Research Council (EP/T022159/, EP/T022159/1)
  • Science and Technology Facilities Council
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene Regulatory Network Analysis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Keywords
gene expression prediction, promoter, deep learning, transcription factor binding motifs, motif co-occurrence, explainable AI, Grad-CAM, in silico mutagenesis, Saccharomyces cerevisiae

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.

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

How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: Predicting gene expression using millions of yeast promoters reveals cis -regulatory logic. https://app.cochat.ai/papers/10.1101/2025.01.25.634853
The paper
Dash, T., & Bornelöv, S. (2025). Predicting gene expression using millions of yeast promoters reveals cis -regulatory logic. Bioinformatics Advances. https://doi.org/10.1101/2025.01.25.634853