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. Previously, we identified the long non-coding RNA NESPR to be specifically expressed in neuroblastoma cells of the adrenergic cell identity. Knockdown of NESPR decreased proliferative potential and ultimately resulted in cell death, highlighting NESPR’s importance in the survival of the adrenergic neuroblastoma cells. Mechanistically, we previously identified NESPR-interacting protein as well as its target genes, both suggesting that NESPR forms R-loop to regulate its target genes. In the research proposal presented here, I tackle several open questions of NESPR’s mechanism of action and how to exploit this mechanistic data to perturb NESPR function. In the first work package, I provide a detailed work flow that aims to decipher in cis regulatory mechanism that NESPR exerts in these adrenergic neuroblastoma cells. The second work package focusses on the implementation of novel methodology to map both RNA-protein and RNA-DNA interaction interfaces. The third and fourth work packages brings together information of all previous work packages to design fully modified antisense oligonucleotides and assess steric blocking of lncRNA-DNA, and –protein interaction interfaces to disrupt NESPR’s function in adrenergic neuroblastoma cells.