You are here

Direct Activation of Nucleobases with Small Molecules for the Conditional Control of Antisense Function

Bardhan A, Brown W, Albright S, Tsang M, Davidson L, Deiters A
Angew Chem Int Ed Engl. 2024 Feb 27:e202318773. doi: 10.1002/anie.202318773. Online ahead of print
Conditionally controlled antisense oligonucleotides provide precise interrogation of gene function at different developmental stages in animal models. Few examples of small molecule-induced activation of antisense function exist, and have been restricted to cyclic morpholinos, which can have significant background activity in the absence of the trigger. Here, we provide a new approach by introducing azido-caged nucleobases that are site-specifically introduced into antisense morpholinos. The caging group design is a simple azidomethylene (Azm) group that, despite its very small size, blocks Watson-Crick base pairing in a programmable fashion. Furthermore, it undergoes facile decaging via Staudinger reduction when exposed to a small molecule phosphine, generating the native antisense oligonucleotide under conditions compatible with biological environments. We demonstrated small molecule-induced gene knockdown in mammalian cells, zebrafish embryos, and Xenopus embryos. We validate the general applicability of this approach by targeting three different genes.
Not Epub
Organism or Cell Type: 
cell culture: HEK293T zebrafish Xenopus laevis
Delivery Method: 
Endo-Porter PEG, microinjection