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DOI: 10.1055/s-0037-1602201
Interfering with the LIN28B-let-7-MYCN pathway in neuroblastoma
Publikationsverlauf
Publikationsdatum:
30. Mai 2017 (online)
Introduction:
LIN28B has been reported to induce neuroblastoma via enhancing MYCN levels through specifically binding precursor let-7 miRNAs and suppressing let-7 miRNA biogenesis. In our experiments we aim to investigate LIN28B inhibition.
Methods:
LIN28B, MYCN and let-7 transcript and protein levels are measured by quantitative RT-PCR and Western blots in different neuroblastoma cell lines. Furthermore, we establish a MYCN-rescue system and luciferase assay in order to monitor inhibition and knockdown of LIN28B by candidate drugs and siRNAs, respectively.
Results and Conclusion:
Expression levels of LIN28B were found to correlate with MYCN expression in neuroblastoma cell lines. We investigate CRISPR/Cas9-mediated knock-out as well as lentivirus-mediated knockdown of LIN28B on let-7 biogenesis in neuroblastoma cell line models. Furthermore, we designed and establish a luciferase- reporter system sensitive to let-7 miRNAs. The combination of those models will provide a platform to monitor the functional activity/relevance of candidate drugs targeting LIN28B.