RESUMEN
Gene and drug delivery systems need crucial update in the issue of nanocarriers. Layered double hydroxides (LDHs) are known as biocompatible inorganic lamellar nanomaterials with versatile properties. In the present study, Zn/Al-LDH nanoparticle was synthesized and characterized by FTIR, XRD, SEM, TEM and Zeta potential tests and then intercalated with valproate and methyldopa by co-precipitation and ion exchange methods. These nanocarriers were applied as high activity nanolayers-based delivery systems. On the other hand, Zn/Al-LDH + plasmid/gene (pCEP4/Cdk9) evaluated on C2C12 myoblast cells. Co-operation loading indicated high efficiency of sorting and release of drugs. Additionally, the Real-Time PCR and Western blotting results for plasmid-gene (pCEP4/Cdk9) delivery showed that Zn/Al-LDH nanoparticles can be used as an effective carrier in cellular uptake and release of genes for gene therapy. Easy and cost-effective production of Zn/Al-LDH nanoparticles proposed them as potential alternatives for the traditional routs of drug/gene delivery.
Asunto(s)
Portadores de Fármacos , Técnicas de Transferencia de Gen , Hidróxidos , Mioblastos/metabolismo , Nanopartículas/química , Animales , Portadores de Fármacos/química , Portadores de Fármacos/farmacología , Hidróxidos/química , Hidróxidos/farmacología , Ratones , Mioblastos/citología , Pentamidina/química , Pentamidina/farmacología , Plásmidos/química , Plásmidos/farmacología , Poliésteres/química , Poliésteres/farmacologíaRESUMEN
Here we developed a complementation method for the study of essential genes in live human cells using the CRISPR/Cas9 system. Proteins encoded by essential genes were expressed using a derivative of the pCEP4 compensating plasmid in combination with Cas9 endonuclease targeting of the chromosomal genes. We show that this strategy can be applied to essential genes, such as those coding for proliferating cell nuclear antigen (PCNA) and DNA polymerase delta subunit 2 (POLD2). As demonstrated for the PCNA protein, our method allows mutational analysis of essential protein-coding sequences in live cells.