Lexogen is the Gold Sponsor of the EMBO | EMBL Symposium “The Complex Life of RNA”

Regulatory small RNAs (sRNAs) play an essential role in mRNA turnover, translational regulation, and chromatin compaction and important regulators of gene expression. These sRNAs associate with specific proteins of the Argonaute family (AGOs) to form RNA-induced silencing complexes (RISCs) and guide the AGO proteins to their respective targets in a process termed RNA interference.

Lexogen’s TraPR Small RNA Isolation Kit greatly facilitates the isolation of RISCs and associated small functional RNAs. It is based on TraPR (Trans-kingdom, rapid, affordable Purification of RISCs), a gel- and bias-free, column-based method (Grentzinger et al., 2020). In a straightforward workflow, freshly harvested or flash frozen tissue or cells are lysed and loaded directly onto TraPR columns. While bulk RNA and DNA are retained on the column, only RISC-associated sRNAs are eluted. These steps are completed in 15 minutes, and the resulting RISC fraction can be analyzed directly or sRNAs can be extracted for further analyses including small RNA sequencing (sRNA-Seq).

Whereas other methods for functional sRNA isolation such as co-immunoprecipitation need species-specific antibodies, TraPR does not require any prior characterization of the sample. Further, by purification of RISCs the TraPR Small RNA Isolation Kit enriches exclusively fully functional, physiologically relevant sRNAs including piRNAs, siRNAs, miRNAs, and scnRNAs. Contaminating RNAs such as degradation products of tRNA, rRNA, and mRNA are effectively excluded from the purified RISC fraction. TraPR is thereby superior to methods that are based on size-selection via columns or gel-purification and cannot distinguish between functional small RNAs and degradation products.

We show, in multiple benchmarking assays, that the TraPR kit considerably improves the quality and consistency of silencing sRNA sample preparation including from notoriously difficult-to-handle tissues/bio-fluids such as starchy storage roots or mammalian plasma, and regardless of RNA contaminants or RNA degradation status of samples. TraPR works consistently over a large input range and eliminates the need for species-specific rRNA depletion (e.g. 2S rRNA in Drosophila).

In summary, the TraPR Small RNA Isolation Kit generates high-quality sRNA preparations suitable for Next Generation Sequencing (NGS) applications and thus provides a highly reproducible, time-saving method that outperforms all current gold-standard procedures for sRNA profiling. Small RNA isolation from liquid samples becomes easily accessible, making TraPR perfectly suited for biomarker discovery applications.

Grentzinger, Thomas; Oberlin, Stefan; Schott, Gregory; Handler, Dominik et al. (2020): A universal method for the rapid isolation of all known classes of functional silencing small RNAs. Nucleic Acids Research. DOI: 10.1093/nar/gkaa472.