Subcellular RNA localization can directly impact the intracellular interactions, morphology, and cellular functions of any given eukaryotic cell. The importance of maintaining proper delivery of mRNA transcripts has previously been studied, with mutations in any component of subcellular localization resulting in abnormal cell developments or functions. Interestingly, mutations in critical cytoskeletal components of microtubules, such as tubulin alpha 1, have been implicated in diseases characterized as “tubulinopathies”, a wide range of brain malfunctions. To gain a deeper understanding of the elements involved in RNA localization and to identify interactions from a multilevel perspective, a combination of techniques were used to identify novel localized RNAs found nearby cytoskeletal structures. To achieve this, we used spatially restricted nucleobase-oxidation to identify novel RNA interactions in close proximity to tubulin, a component of microtubules. In this technique, a localized light reactive fluorophore can be used to produce oxygen radicals with the ability to tag RNA transcripts within a short diffusion radius, thus, allowing for the extraction and identification of RNA and their interactions. As a means for further understanding tubulinopathies and novel RNA interactions within tubulin, fluorophore J646 was localized to tubulin alpha 1 and microtubule-acting cross-linking factor 43 proteins and used to tag nearby RNA. To verify these novel RNA interactions, single-molecule fluorescence in Situ hybridization (smFISH), a technique that allows for the study of localized RNA, DNA, or protein, was to be used. Although this technique allows for visualization of in vivo RNA localization, it involves the production of expensive fluorescently labeled oligonucleotide probes that may not possess high sensitivity to detect RNA in low abundance. Therefore, to verify novel RNA interactions utilizing microtubules for localization with increased detection, a procedure for generating smFISH probes from readily available oligonucleotides with more than one attached dye was created. Although further optimizations are required before usage, I have successfully attached more than one dye to the unlabeled smFISH probes. By pairing both nucleobase oxidation and smFISH, a greater understanding of novel RNA localization utilizing microtubules can be achieved. Furthermore, by developing a protocol to generate highly sensitive smFISH probes by increasing the attached dye counts from one to several, a greater fluorescence can be achieved allowing for higher sensitivity of even the smallest quantities of localized RNA.