RiboMarker ® Platform

RiboMarkers are highly informative markers of disease and disease progression for conditions ranging from cancer to infectious diseases like funguses or viruses such as COVID. RiboMarkers can be used to determine not only whether a patient has a disease but also the state of the disease’s progression, for example, dissemination of an infection from the primary site throughout the body, markers of residual disease or possibly even of disease termination.

Our first RiboMarker diagnostic is for the fungal disease known as Valley Fever (VF). VF is a debilitating disease most often found in the Western states such as Arizona and CA and in Mexico and South America. When VF disseminates throughout the body patients require lifelong antifungal treatments that are costly and not very effective. The VF RiboMarker program allows better detection and analysis of dissemination and residual disease in these patients. VF and other “dimorphic” funguses diseases such as Histoplasmosis are dramatically increasing due to climate change.

Scientists at RealSeq are committed to expanding the RiboMarker diagnostic platform to address other fungal diseases as well as any other condition where the ability to accurately predict, identify, and track dissemination critically impacts patient outcomes.

RealSeq® technology for small RNA sequencing library preparation

Accuracy through circularization

RealSeq Biosciences’ core expertise includes innovative proprietary technologies for bias-free small RNA/miRNA NGS library construction, targeted NGS tools, and cf-RNA analysis (liquid biopsy) that form the basis of the Company’s life science research programs and product development.

The Company is the recipient of several NIH SBIR Phase I/II grants including fundng for its RealSeq NGS library construction platform.

RealSeq® is a novel method for preparing small-RNA sequencing libraries that nearly eliminates incorporation bias in Next-Generation-Sequencing (NSG). This technology solves the problem of commonly used sequencing library preparations that lead to underdetection of many miRNAs, some by as much as 10,000-fold.

Under-representation in sequencing libraries can obscure the presence of some RNAs, including potential biomarkers. Accurate quantification of microRNAs (miRNA) and other small RNAs is important for understanding their biology and for developing new biomarkers and therapeutic targets.

Most bias stems from sequence-dependent variability in the enzymatic ligation reactions that attach the two adapters to the 3’ and 5’ ends of the miRNAs /small RNAs during preparation of sequencing libraries. By using a novel single adapter and circularization, RealSeq® greatly reduces library preparation bias.

See our publication: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6120088/

Sergio Barberán-Soler* , Jenny M. Vo, Ryan E. Hogans, Anne Dallas, Brian H. Johnston and Sergei A. Kazakov* (2018) Decreasing miRNA sequencing bias using a single adapter and circularization approach. Genome Biology, 19:105.

Targeted Approach

RealSeq-T is a novel technology that allows the targeted, direct quantification of microRNAs from biofluid samples. Data from plasma samples show that employing a targeted sequencing approach eliminates the need to perform RNA extraction and is significantly more reproducible compared to non-targeted approaches that detect microRNAs after standard RNA extraction.

Single Cell

RealSeq®-SC is an innovative and highly efficient technology to improve the detection accuracy of single-cell small RNA sequencing and minimize technical dropout rates. The technology expands single-cell transcriptomics to encompass small as well as large RNAs, is effective across multiple cell types, and provides an effective platform for single cell biomarker/liquid biopsy applications.

Selected Suppression Probes - SSP

Highly-expressed RNA species reduce read depth of less abundant species that researchers care about. RealSeq Biosciences has developed a targeted removal probe (Selective Suppression Probe- SSP™) approach to depleting over-abundant species from RNA samples, resulting in increased miRNA reads and greater sequencing depth of miRNAs.