To date, OTTR has been used for quantifying tRNA species. This method provides a reliable and precise quantification of tRNAs and allows for the detection of changes in tRNA levels under different physiological conditions. Therefore, this method is useful for a variety of research applications in the field of molecular biology and genetics.
As tRNAs are essential components of translation, the ability to quantify tRNA levels accurately is crucial for understanding how the translation machinery is regulated. Using OTTR, researchers can determine changes in tRNA levels in response to different growth conditions, environmental stress, or genetic modifications. This information can help to identify factors that affect tRNA abundance and their potential roles in modulating translation.
In particular, the OTTR protocol has been used to characterize the small RNA composition of mammalian sperm populations. This work revealed that sperm small RNA pools are composed largely of rRNA fragments and both 5 and 3 tRNA halves from the majority of tRNAs. This improved understanding of sperm payload composition has implications for our understanding of the biogenesis of structural RNA fragments in the male germline, as well as the biochemical nature of the RNAs delivered to the zygote upon fertilization.
The OTTR protocol could also be used in the study of piwi-interacting RNAs (piRNAs), which is another important classes of small RNAs. While the applications of OTTR have mainly been focused on tRNA fragment detection, OTTR has also been shown to perform well in the capture of miRNAs which could be useful for the study of miRNA expression patterns in different cell types or under different conditions.
Future applications of protocols similar to OTTR are also being considered in the development of diagnostic assays for small RNAs. The high fidelity of the OTTR protocol in capturing small RNAs could make it an attractive option when paired with liquid biopsy assays, where circulating small RNAs are analyzed as biomarkers for various diseases.