Journal article
Cationic nanocapsules as a non-viral vector for CRISPR-Cas9–based genome editing for myocilin associated glaucoma therapy
AIMS molecular science, Vol.12(4), pp.341-356
2025
DOI: 10.3934/molsci.2025020
Abstract
Our aim of this work was the development of positively charged D-α-tocopheryl glycol succinate 1000 (TPGS)-chitosan nanocapsules (TCNs) as a non-viral vector for effective genome editing. A two-step process was used to create cationic TCNs. TPGS was initially esterified using succinic anhydride to add a carboxylic acid group. Then, using a complex coacervation technique, ionic cross-linking was used to join the activated TPGS with chitosan. Fourier transform infrared (FTIR) spectroscopy was used to validate the synthesis of TCNs. A gel retardation experiment was performed to evaluate TCNs/pDNA complex entrapments at various N/P ratios (2:1, 5:1, and 10:1). The physicochemical properties of the TCNs/pDNA complex were identified. Plasmid DNA was protected from DNase I activity through TCN complexation. HEK293 and GTM3 cell lines were used to test in vitro transfection efficiency. At N/P 5:1, TCNs/pDNA exhibited the maximum transfection efficiency, which was comparable to Lipofectamine 3000™. Cellular uptake and localization studies confirmed effective delivery of Cas9-GFP plasmids using TCNs. A T7 endonuclease mismatch detection assay further demonstrated effective targeting and editing of the MYOC gene in HEK293 cells using our TCN delivery system. Based on these findings, cationic TCNs represent a promising non-viral gene delivery platform for potential treatment of myocilin-associated glaucoma.
Details
- Title: Subtitle
- Cationic nanocapsules as a non-viral vector for CRISPR-Cas9–based genome editing for myocilin associated glaucoma therapy
- Creators
- Karthikeyan KesavanVirendra KumarJothimani RajeswariCharles SearbyVal C. Sheffield
- Resource Type
- Journal article
- Publication Details
- AIMS molecular science, Vol.12(4), pp.341-356
- DOI
- 10.3934/molsci.2025020
- ISSN
- 2372-0301
- Number of pages
- 16
- Language
- English
- Date published
- 2025
- Academic Unit
- Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Medical Genetics and Genomics; Ophthalmology and Visual Sciences
- Record Identifier
- 9985019042802771
Metrics
1 Record Views