Assessment of CRISPRa-mediated gdnf overexpression in an In vitro Parkinson's disease model

体外帕金森病模型中 CRISPRa 介导的 gdnf 过度表达的评估

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作者:Paula Guzmán-Sastoque, Sebastián Sotelo, Natalia P Esmeral, Sonia Luz Albarracín, Jhon-Jairo Sutachan, Luis H Reyes, Carolina Muñoz-Camargo, Juan C Cruz, Natasha I Bloch

Discussion

The delivery system exhibits significant endosomal escape of more than 56%, crucial for the effective delivery and activation of the genetic material within cells. The increased gdnf expression correlates with a notable reduction in MAO-B complex activity, reaching basal values of 14.8 μU/μg of protein, and a reduction in reactive oxygen species. Additionally, there is up to a 34.6% increase in cell viability in an In vitro Parkinson's disease model treated with the neurotoxin MPTP. Our study shows that increasing gdnf expression can remediate some of the cellular symptoms associated with Parkinson's disease in an in vitro model of the disease using a novel nanostructured delivery system.

Methods

We have developed a targeted delivery system using a magnetite nanostructured vehicle for the efficient transport of genetic material. This system has resulted in a substantial increase, up to 200-fold) in gdnf expression in an In vitro model of Parkinson's disease using a mixed primary culture of astrocytes, neurons, and microglia.

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