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ORIGINAL RESEARCH article

Front. Bioeng. Biotechnol.
Sec. Nanobiotechnology
Volume 12 - 2024 | doi: 10.3389/fbioe.2024.1420183

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

Provisionally accepted
Paula Guzmán-Sastoque Paula Guzmán-Sastoque 1Sebastián Sotelo Sebastián Sotelo 1Natalia P. Esmeral Natalia P. Esmeral 1Sonia L. Albarracin Sonia L. Albarracin 2Jhon J. Sutachan Jhon J. Sutachan 2Luis H. Reyes Luis H. Reyes 3Carolina Muñoz-Camargo Carolina Muñoz-Camargo 1Juan C. Cruz Juan C. Cruz 1,3*Natasha I. Bloch Natasha I. Bloch 1*
  • 1 Department of Biomedical Engineering, Faculty of Engineering, University of Los Andes, Bogotá, Cundinamarca, Colombia
  • 2 Department of Nutrition and Biochemistry, Faculty of Sciences, Pontifical Javeriana University, Bogota, Colombia
  • 3 Department of Chemical and Food Engineering, University of Los Andes, Colombia, Bogotá, Cundinamarca, Colombia

The final, formatted version of the article will be published soon.

    Parkinson's disease (PD) presents a significant challenge in medical science, as current treatments are limited to symptom management and often carry significant side effects. Our study introduces an innovative approach to evaluate the effects of gdnf overexpression mediated by CRISPRa in an in vitro model of Parkinson's disease. The expression of gdnf can have neuroprotective effects, being related to the modulation of neuroinflammation and pathways associated with cell survival, differentiation, and growth. We have developed a targeted delivery system utilizing a magnetite nanostructured vehicle for the efficient transport of genetic material, resulting in a substantial increase (up to 200-fold) in gdnf expression in a mixed primary culture of astrocytes, neurons, and microglia. This delivery system exhibits significant endosomal escape of more than 56%, crucial for the effective delivery and activation of the genetic material within cells. Our findings demonstrate that the increased gdnf expression correlates with a notable reduction in MAO-B complex activity, reaching basal values of 14.8 μU/μg of protein, an 80% decrease in reactive oxygen species, and 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 increasing gdnf expression can remediate some of the cellular symptoms associated with Parkinson's disease an in vitro model of the disease using a novel nanostructured delivery system.

    Keywords: CRISPR gene overexpression, Parkinson's disease model, gdnf, Nanoparticle delivery system, Oxidative Stress

    Received: 19 Apr 2024; Accepted: 25 Jul 2024.

    Copyright: © 2024 Guzmán-Sastoque, Sotelo, Esmeral, Albarracin, Sutachan, Reyes, Muñoz-Camargo, Cruz and Bloch. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Juan C. Cruz, Department of Chemical and Food Engineering, University of Los Andes, Colombia, Bogotá, 111711, Cundinamarca, Colombia
    Natasha I. Bloch, Department of Biomedical Engineering, Faculty of Engineering, University of Los Andes, Bogotá, Cundinamarca, Colombia

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.