Neuroblastoma is the most common extracranial solid tumour of early childhood and accounts for 15% of paediatric cancer deaths, despite accounting for only 8% of cases. Understanding the molecular mechanisms that contribute to aggressive neuroblastoma will provide a platform for the development of more potent and specific treatments for this malignancy. Past studies have shown that activity of the immortalising enzyme telomerase and expression of telomerase reverse transcriptase (hTERT) in primary neuroblastoma samples correlates with patient outcome. However, our analysis of a publically available microarray dataset showed that expression of the gene encoding the telomerase RNA-binding protein, dyskerin (DKC1) was more highly prognostic of event-free survival than hTERT expression. Multivariate analysis showed that DKC1 expression was independent of established prognostic indicators such as INSS stage and MYCN amplification. In a panel of 15 neuroblastoma cell lines, dyskerin gene and protein expression was found to be high irrespective of MYCN amplification. The functional significance of dyskerin levels in neuroblastoma was investigated by siRNA-mediated repression of dyskerin and/or stable suppression of dyskerin using shRNA. Dyskerin repression caused a concomitant reduction in the level of the telomerase RNA component (hTR), but had no effect on hTERT and variable effects on telomerase enzyme activity. Repression of dyskerin compromised cell viability, impaired proliferation and inhibited anchorage-independent growth of neuroblastoma cells, including the putative telomerase-negative cell line SK-N-FI. Reduced expression of dyskerin in a neuroblastoma cell line without MYCN amplification potentiated the effects of the chemotherapeutic agents Cisplatin and Doxorubicin, and the mTOR inhibitor, Rapamycin. Taken together, these data demonstrate that high levels of dyskerin contribute to the malignant properties of neuroblastoma cells and support further investigation of dyskerin as a therapeutic target in the treatment of refractory neuroblastoma.