The current technological landscape of gene detection is largely reliant on slow and overly complicated methods of determining the presence of particular genes in a sample. Ultrafast gene detection (UFGD) via barcoded dead Cas9 proteins (dCas9) aims to reform gene detection, making it highly portable, accessible and most importantly fast. PolyC9, made from 22 units of C9, is a soluble proteinaceous pore that exhibits many characteristics which make it an ideal candidate for use in this novel nanopore sensing technology. C9 protein is easily produced in mammalian expression cultures, readily forms soluble pores that do not require extraction or additional solubilisation steps, is stable within a MinION flow cell (a key technology in the Oxford Nanopore Technologies, ONT, platform) and has a large β-barrel. However, the lumen is restricted by 3 rings of N-glycans that reduce the functional lumen diameter to ~ 10 nm, which is hypothesised to be too narrow to passage a dCas9 through its lumen. This issue was in part mitigated by removing the N-glycans, thereby unrestricting dCas9 passage. However, glycosylation is required for secretion of mammalian proteins such as C9, and this aglycosylation resulted in poor secretion and reduced yield. Non-native N-glycans were introduced outside of the PolyC9 lumen in order to ameliorate the poor secretion of the completely aglycosylated C9 mutant. Secretion of novel N-glycan mutants was assessed by western blot analysis, and pore stability observed via negative stain electron microscopy. Encouragingly, three out of five mutations resulted in improved secretion. These successful mutants were assessed by scientists at ONT for changes in their electrophysiology properties, and remarkably 3 mutants exhibited increased maximum voltages, indicative of a less restricted pore lumen. These results represent acritical step towards UFGD biosensors and showcase the impact N-glycans can have in protein engineering.