The human genome comprises approximately six billion DNA nucleotides distributed across 23 pairs of chromosomes, encoding the biological processes underlying health and disease.
As genomic science accelerates, the tools for probing gene function must evolve to match this. At Revvity’s Dharmacon™ reagents team, we develop tools that enable precise genome interrogation and modulation to support functional genomics, target validation, and translational research. Central to this effort is the accuracy and currency of the reference data used to design genomic reagents.
Dharmacon’s siRNA designs integrate curated RefSeq annotations from the National Center for Biotechnology Information (NCBI) with a proprietary Dharmacon design algorithm1. Because RefSeq defines gene structures and transcript boundaries, it serves as a critical data source for selecting RNAi designs that specifically target the gene of interest. As transcript annotations continue to evolve, aligning siRNA designs with the latest bioinformatic and biological knowledge ensures sustained accuracy, confidence, and reproducibility.
The evolving transcriptome and why RefSeq alignment matters
Advances in sequencing technologies and data curation have dramatically expanded RefSeq to approximately 175,000 human transcripts, uncovering increased isoform complexity, regulatory nuances, and refined transcript boundaries across protein-coding genes. ON-TARGETplus™ 2.0 leverages these advances by directly incorporating the latest RefSeq annotations into siRNA design, ensuring reagents target the most current and biologically relevant transcript definitions (Figure 1). As transcript annotations continue to evolve whether by refining canonical records, elevating alternative isoforms, or redefining gene boundaries based on new evidence; ON TARGETplus 2.0 siRNAs stay aligned with the latest RefSeq to maintain precise, up to date targeting. This alignment supports superior transcript coverage and dependable target engagement, resulting in more consistent knockdown, clearer phenotypes, and highly reproducible outcomes, particularly in large scale screens and hit validation workflows.
Figure 1- Depicts the expansion of transcript isoforms enabled by advances in sequencing technology and illustrates how ON TARGETplus 2.0 siRNA remains aligned to the updated RefSeq to ensure accurate, current targeting.
Quick facts – ON-TARGETplus 2.0
- What is the difference between the legacy ON-TARGETplus reagents and ON-TARGETplus 2.0?
- Is there a price premium on ON-TARGETplus 2.0?
No. ON-TARGETplus 2.0 is priced in line with legacy ON-TARGETplus. Customers get additional benefits at the same cost.
A: ON-TARGETplus 2.0 is a next-generation upgrade of the original ON-TARGETplus siRNA. It features improved design algorithms, continuous transcriptome realignment, and updated sequence ranking to enhance specificity, minimize off-target effects, and ensure consistent gene knockdown. In contrast, legacy ON-TARGETplus designs were based on earlier transcriptome data and do not include these ongoing optimizations.