Nucleic acid therapeutics is an emerging and rapidly advancing field with the potential to target previously undruggable genes. These therapeutics include antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), circular RNA (circRNA), large RNAs like messenger RNA (mRNA), and other emerging technologies. These therapies can potentially manipulate gene expression and produce therapeutic proteins, making them suitable for various pathologies with well-defined genetic targets, including infectious diseases, cancers, immune disorders, and Mendelian disorders, including neurological conditions.

Each therapeutic uses different molecular mechanisms to achieve gene knockdown, gene upregulation, gene expression/replacement, and gene editing. Recent advancements in genome sequencing, single-cell gene expression analysis, and gene manipulation techniques have expanded the discovery of new targets for gene therapies. However, effectively manipulating these targets requires efficient delivery of therapeutic RNA to diseased cells. Thus, the discovery and development of these therapies involve not only molecular biology techniques but also delivery biology and engineering.

Chemical modifications, bioconjugation to receptor-targeted ligands, or encapsulation in lipid or nucleotide nanocarriers enhance the delivery of oligonucleotide therapeutics. A range of different assays are utilized to design and test the delivery strategies mentioned above. These include surface proteomics, reporter-based assays for testing endosomal escape, SELEX for aptamer discovery, microscopy for nanocarrier characterization, gene knockdown assays, Ago2 loading assays, and novel nucleotide analytical assays, among others.

This Methods Collection aims to bring together a diverse set of wet-lab and dry-lab methods that aid in building nucleic acid therapeutic platforms.