Evaluating stably expressed genes in single cells
Abstract
Background
Single-cell RNA-seq (scRNA-seq) profiling has revealed remarkable variation in transcription, suggesting that expression of many genes at the single-cell level are intrinsically stochastic and noisy. Yet, on cell population level, a subset of genes traditionally referred to as housekeeping genes (HKGs) are found to be stably expressed in different cell and tissue types. It is therefore critical to question whether stably expressed genes (SEGs) can be identified on the single-cell level, and if so, how their expression stability can be assessed? We have developed a computational framework for ranking expression stability of genes in single cells. Here we evaluate the proposed framework and characterize SEGs derived from two scRNA-seq datasets that profile early human and mouse development.
Results
Here, we show that gene expression stability indices derived from the early human and mouse development scRNA-seq datasets are highly reproducible and conserved across species. We demonstrate that SEGs identified from single cells based on their stability indices are considerably more stable than HKGs defined previously from cell populations across 10 diverse biological systems. Our analyses indicate that SEGs are inherently more stable at the single-cell level and their characteristics reminiscent of HKGs, suggesting their potential role in sustaining essential functions in individual cells.
Conclusions
SEGs identified in this study have immediate utility both for understanding variation/stability of single-cell transcriptomes and for practical applications including scRNA-seq data normalization, the proposed framework can be applied to identify genes with stable expression in other scRNA-seq datasets.
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