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bioRxiv · 10.1101/2020.10.20.346908

Resolving a Systematic Error in STARR-seq for QuantitativeEnhancer Activity Mapping

Hou, C.·Niu, L.·Wan, J.·Sun, J.·Huang, Y.·He, N.·Li, L.
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Abstract

STARR-seq assesses millions of fragments in parallel measuring enhancer activity quantitatively. Here we show that STARR-seq is critically flawed with a systematic error in the cells of Arabidopsis thaliana (A. thaliana). Large amount of self-transcripts (STs) is lost during reverse transcription because these STs are polyadenylated after alternative polyadenylation sites (APAS) inside the test sequences. We solved this problem by using specially designed primer and recovered self-transcribed sequences independent from the PAS usage. In A. thaliana, we identified active enhancers and also enhancers quiescent in their endogenous genomic loci. Different from traditional STARR-seq identified enhancers, enhancers identified by new method are highly enriched in sequences proximal to the 5 and 3 ends of genes, and their epigenetic states correlate with gene expression levels. Our solution applies to methods based on self-transcript quantification. In addition, our results provide an invaluable functional enhancer activity map and insights into the functional complexity of enhancers in A. thaliana.

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Hou, C., Niu, L., Wan, J., Sun, J., Huang, Y., He, N., Li, L.. 2020-10-21. Resolving a Systematic Error in STARR-seq for QuantitativeEnhancer Activity Mapping. https://doi.org/10.1101/2020.10.20.346908

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