In this study, direct and effective intracellular delivery of CRISPR/Cas9 plasmids for homology‐directed repair is achieved by functionalized mesoporous silica nanoparticles (MSNs). The functionalized MSNs (Cy5.5‐MSNs‐NLS) are synthesized by in situ labeling of a fluorescent dye (Cy5.5) and surface conjugation of nuclear localization sequence (NLS, PKKKRKV), showing a high loading efficiency (50%) toward the plasmids (PXN cutdown plasmid: GFP‐Cas9‐paxillin_gRNA and repair plasmid: AICSDP‐1: PXN‐EGFP). Subsequently, a polymeric coating of the poly(dimethyldiallylammonium chloride) (PDDA) is electrostatically deposited onto the plasmid‐loaded Cy5.5‐MSNs‐NLS by microfluidic nanoprecipitation. The coating layer offers effective protection against the denaturation of plasmids by EcoRV restriction enzymes, and is shown to prevent premature release. Moreover, owing to the positive charge and pH‐responsive disaggregation of PDDA, enhanced cellular internalization (16 h) and endosomal escape (4 h) of the nanocarrier are observed. After escape of nanocarrier system into the cytoplasm, the NLS on the surface of MSNs facilitates nuclear transport of the CRISPR/Cas9 plasmids, achieving successful GFP‐tag knock‐in of the PXN genomic sequence in U2OS cells. This intracellular delivery system thus offers an attractive method to overcome physiological barriers for CRISPR/Cas9 delivery, showing considerable promise for paxillin‐associated focal adhesion and signaling regulator investigation.
Appropriate tuning of robust artificial coatings can not only enhance intracellular delivery, but also preserve the biological functions of genetic molecules in gene based therapies. Here, we report a strategy...
Triple‐negative breast cancer (TNBC) is still the most aggressive cancer in women. Combination chemotherapy holds great potential for cancer therapy; however, the off‐target and side effects of free chemotherapy administration remain a major challenge. In this study, we developed a photo/thermo‐responsive nanoplatform that can be used for TNBC treatment via photothermic therapy in combination with multidrug therapy. By conjugating the chemotherapy drug PTX prodrug on the surface of mesoporous silica‐coated gold nanorod nanoparticles and then loading another chemotherapy drug, CPT, the Au@MSN‐PTX@CPT nanoparticles exhibited great photothermal response, redox response drug release and cancer cell inhibition abilities. Otherwise, we further coated the Au@MSN‐PTX@CPT nanoparticle with a temperature‐sensitive polymer poly(N‐isopropylacrylamide‐co‐methacrylic acid) (p(NIPAM‐co‐MAAc)), and the polymer‐coated Au@MSN‐PTX@TPT@polymer nanoparticles showed perfect near‐infrared (NIR) light controlled drug release. Finally, the Au@MSN‐PTX@CPT@polymer nanoparticles were injected into the 4T1 breast cancer mouse model. The Au@MSN‐PTX@CPT@polymer nanoparticles preferably accumulated at the tumor site and had reduced chemotherapy injuries and great antitumor activity when combined with 650 nm laser treatment. In summary, our developed Au@MSN‐PTX@CPT@polymer nanoparticles served as a good method for controlled chemodrug delivery and provided a good choice for TNBC combination therapy.
Ribonucleoprotein (RNP) based CRISPR/Cas9 gene-editing system shows great potential in biomedical applications. However, due to the large size, charged surface and high biological sensitivity of RNP, its efficient delivery with...
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