Lipophilic Conjugates for Carrier-Free Delivery of RNA Importable into Human Mitochondria

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Dovydenko, Ilya | Meschaninova, Mariya | Heckel, Anne-Marie | Tarassov, Ivan | Venyaminova, Alya | Entelis, Nina

Edité par CCSD -

International audience. Defects in human mitochondrial genome can cause a wide range of clinical disorders which still do not have efficient therapies. The natural pathway of small non-coding RNA import can be exploited to address into mitochondria therapeutic RNAs. To create an approach of carrier-free targeting of RNA into living human cells, we designed conjugates containing a cholesterol residue and developed the protocols of chemical synthesis of oligoribonucleotides conjugated with cholesterol residue through cleavable pH-triggered hydrazone bond. The biodegradable conjugates of importable RNA with cholesterol can be internalized by cells in a carrier-free manner; RNA can then be released in the late endosomes due to a change in pH and partially targeted into mitochondria. Here we provide detailed protocols for solid-phase and "in solution" chemical synthesis of oligoribonucleotides conjugated to a cholesterol residue through a hydrazone bond. We describe the optimization of the carrier-free cell transfection with these conjugated RNA molecules and methods for evaluating the cellular and mitochondrial uptake of lipophilic conjugates.

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