Long non-coding RNAs (lncRNAs) are a class of transcripts with lengths exceeding 200 nucleotides that do not encode proteins. Despite their crucial roles in cellular functions and biological processes, only a minority of the over 20,000 annotated lncRNAs have been functionally characterized. Here, we established a high-throughput, CRISPR-Interference (CRISPRi) arrayed screening plaKorm with serial cellular and molecular phenotyping to systematically characterize lncRNA functions. We reasoned that the integration of a comprehensive cellular and molecular phenotype can increase the probability of uncovering cellular functions and pathways controlled by lncRNA transcripts.
Neuroblastoma is a childhood cancer of the sympathetic nervous system. Recent studies have shown that neuroblastoma tumors are composed of two cell identities, i.e. the adrenergic and mesenchymal identity. Both identities are driven by a core regulatory transcriptional circuitry, which acts as an autoregulatory positive feedforward loop, to delineate the cell identity through regulation of its target genes. We identified the long non-coding RNA NESPR to be specifically expressed in neuroblastoma cells of the adrenergic cell identity.