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Two-Photon-Induced Charge-Variable Conjugated Polyelectrolyte Brushes for Effective Gene Silencing.

Cationic conjugated polyelectrolytes can absorb negatively charged small interfering RNA (siRNA) and also visualize the cellular internalization of siRNA, which thus have been extensively explored as siRNA carriers. However, their low charge density cannot afford a high carrying capability, severely impeding gene transfection efficiency. Moreover, the intracellular controlled release of siRNA is another factor that limits the widespread use of siRNA therapeutics. Herein, we present a novel two-photon-induced charge-variable conjugated polyelectrolyte brush as an efficient siRNA carrier. This cationic conjugated polyelectrolyte brush (PPENBr-ONB) with densely cationic charges produces remarkable carrying capability with siRNA. In addition, PPENBr-ONB with large two-photon absorption (TPA) cross-section represents effective fluorescence resonance energy transfer (FRET) to photoresponsive side chain with 720 nm illumination for two-photon-induced photolysis. Hence, the charge transformation of the photoresponsive side chain from cations to zwitterions would remarkably elevate siRNA release. The obtained PPENBr-ONB shows considerable fluorescence quantum yields (0.16) in aqueous solution, sufficient to serve as a reporter for cellular imaging. Agarose gel electrophoresis experiments indicate that PPENBr-ONB exhibit excellent siRNA-loading capacity (1 mol PPENBr-ONB to more than 20 mol siRNA). Furthermore, PPENBr-ONB with large TPA cross-section (1.47 × 105 GM) exhibits promoted siRNA release (78%) under 720 nm illumination. In vitro experiment shows that PPENBr-ONB/siRNA complex could efficaciously knock out of targeted Plk1 mRNA to 24.7% under 720 nm illumination for 1 h. This two-photon excitation siRNA carrier offers an efficacious strategy for the exploitation of photo controlled gene delivery system.