SpliceRead: Improving Canonical and Non-Canonical Splice Site Prediction with Residual Blocks and Synthetic Data Augmentation

Abstract

Accurate splice site prediction is fundamental to understanding gene expression and its associated disorders. However, most existing models are biased toward frequent canonical sites, limiting their ability to detect rare but biologically important non-canonical variants. These models often rely heavily on large, imbalanced datasets that fail to capture the sequence diversity of non-canonical sites, leading to high false-negative rates. Here, we present SpliceRead, a novel deep learning model designed to improve the classification of both canonical and non-canonical splice sites using a combination of residual convolutional blocks and synthetic data augmentation. SpliceRead employs a data augmentation method to generate diverse non-canonical sequences and uses residual connections to enhance gradient flow and capture subtle genomic features. Trained and tested on a multi-species dataset of 400- and 600-nucleotide sequences, SpliceRead consistently outperforms state-of-the-art models across all key metrics, including F1-score, accuracy, precision, and recall. Notably, it achieves a substantially lower non-canonical misclassification rate than baseline methods. Extensive evaluations, including cross-validation, cross-species testing, and input-length generalization, confirm its robustness and adaptability. SpliceRead offers a powerful, generalizable framework for splice site prediction, particularly in challenging, low-frequency sequence scenarios, and paves the way for more accurate gene annotation in both model and non-model organisms.The open sourced code of SpliceRead and a detailed documentation is available at The open-sourced code of SpliceRead and detailed documentation are available at https://github.com/OluwadareLab/SpliceRead.