Various off-target mRNAs with partial complementarity to the ADAM17 shRNA guide strand (ADAM17 shRNA off-targets)

This entry refers to the unintended silencing of various messenger RNAs (mRNAs) that possess partial sequence complementarity to the guide strand of a short hairpin RNA (shRNA) designed to target ADAM Metallopeptidase Domain 17 (ADAM17). Such off-target effects are a major challenge in RNA interference (RNAi) technology, occurring when the "seed region" of the shRNA (nucleotides 2-8) binds to the 3' untranslated regions (UTRs) of non-target transcripts (Jackson et al., 2003). This binding mimics the natural microRNA (miRNA) pathway, leading to the degradation or translational repression of genes unrelated to the intended target (Birmingham et al., 2006). ADAM17, also known as TACE (TNF-alpha converting enzyme), is a membrane-bound metalloproteinase responsible for the proteolytic cleavage of various cell surface proteins, including TNF-alpha and EGFR ligands (Moss et al., 1997). When shRNAs targeting ADAM17 are used, the unintended knockdown of off-target mRNAs can result in significant cellular toxicity or misleading phenotypic changes (Grimm et al., 2006). These off-target interactions are particularly problematic in therapeutic development, as they can trigger innate immune responses or cause systemic organ damage, such as hepatotoxicity. To mitigate these risks, researchers employ advanced bioinformatic algorithms to predict off-target binding and utilize chemical modifications to the shRNA backbone to enhance specificity. Understanding the profile of these off-target mRNAs is essential for the safety assessment of any RNAi-based drug candidate targeting the ADAM17 pathway.

Other names

RNAi off-target effectsSeed-mediated off-target silencingmiRNA-like off-target effectsNon-specific mRNA knockdownADAM17 shRNA-induced off-targets

Seed-sequence-mediated mRNA degradation or translational repression via the RNA-induced silencing complex (RISC) (Jackson et al., 2003).