Open-access synthetic spike-in mRNA-seq data for cancer gene fusions

Oncogenic fusion genes underlie the mechanism of several common cancers. Next-generation sequencing based RNA-seq analyses have revealed an increasing number of recurrent fusions in a variety of cancers. However, absence of a publicly available gene-fusion focused RNA-seq data impedes comparative assessment and collaborative development of novel gene fusions detection algorithms. Scientists at Translational Genomics Research Institute and Illumina have generated nine synthetic poly-adenylated RNA transcripts that correspond to previously reported oncogenic gene fusions. These synthetic RNAs were spiked at known molarity over a wide range into total RNA prior to construction of next-generation sequencing mRNA libraries to generate RNA-seq data.

Leveraging a priori knowledge about replicates and molarity of each synthetic fusion transcript, the scientists demonstrate utility of this dataset to compare multiple gene fusion algorithms’ detection ability. In general, more fusions are detected at higher molarity, indicating that our constructs performed as expected. However, systematic detection differences are observed based on molarity or algorithm-specific characteristics. Fusion-sequence specific detection differences indicate that for applications where specific sequences are being investigated, additional constructs may be added to provide quantitative data that is specific for the sequence of interest.


Tembe WD, Pond SJ, Legendre C, Chuang HY, Liang WS, Kim NE, Montel V, Wong S, McDaniel TK, Craig DW, Carpten JD. (2014) Open-access synthetic spike-in mRNA-seq data for cancer gene fusions. BMC Genomics 15(1):824. [abstract]


Translational Genomics Research Institute