Expanded Gene Targeting in RNA Hacking With G‐Tract‐Supply Staple Oligomers

ABSTRACT RNA hacking (RNAh) is a gene regulation technology that employs a short oligonucleotide, termed a Staple oligomer, to induce the formation of RNA G‐quadruplex structures on target mRNAs. While RNAh has the potential to target approximately 65% of human mRNAs, its applicability to the remaining genes is restricted by the sequence constraints. Herein, we present the G‐tract‐supply Staple oligomer (Gs‐Staple oligomer), designed to expand the range of targetable mRNAs within the RNAh framework. Incorporating G‐tracts into Staple oligomers alleviates the sequence constraints, enabling access to a broader range of mRNA targets. Gs‐Staple oligomers effectively suppressed the translation of target proteins in mammalian cells and in vivo. Furthermore, the gene suppression could be precisely modulated by adjusting the linker length between the G‐tracts. These findings have significantly expanded the versatility of RNAh, suggesting its potential for further development while highlighting its potential to be utilized as a nucleic acid‐based tool for research and clinical medicine.