Telomere maintenance is essential for the continued proliferation of mitotically active cells. Approximately 85-90% of cancers ensure replicative immortality by activating the enzyme telomerase, a ribonucleoprotein complex consisting of the catalytic reverse transcriptase hTERT and the RNA template component hTR. Telomere maintenance via telomerase requires proper assembly of its protein and RNA components, as well as the recruitment of cofactors involved in maturation, stability, and telomere recruitment. The DBHS family of RNA/DNA binding proteins (NONO, SFPQ, and PSPC1) are involved in several aspects of RNA processing and DNA repair. We have investigated the effects of all three DBHS proteins on aspects of telomerase assembly and recruitment to the telomere. RNA immunoprecipitation revealed that all three DBHS proteins interact with hTR in telomerase positive cancer cell lines. DBHS protein depletion significantly reduced hTR recruitment to the telomere, with NONO and SFPQ depletion causing hTR sequestration into large nuclear foci identified as Cajal bodies. Telomerase activity, measured by a direct telomerase activity method, revealed that DBHS protein depletion decreased telomerase catalytic activity by preventing hTERT association with hTR, indicating that these proteins play a role in the assembly of the catalytic core of telomerase. All three DBHS proteins also co-purified with hTERT and did not significantly alter telomerase expression, suggesting direct roles in regulating catalytic activity. Finally, CRISPR-Cas9 knockout of NONO resulted in progressive telomere shortening in clonal populations of telomerase positive cancer cells. Our results identify the DBHS proteins as novel and unexpected players in the regulation of telomerase recruitment/activity that have the potential to be exploited to remove cancer cell immortality and prevent/inhibit secondary tumour immortalisation.