July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Characterization of magnetofection transfection reagent, a novel transfection method for gene therapy in a mouse model
Author Affiliations & Notes
  • Priyanka Priyadarshani
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Salma Ferdous
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Shanu Markand
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Kevin Joseph Donaldson
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Jeffrey H Boatright
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • J M Nickerson
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Footnotes
    Commercial Relationships   Priyanka Priyadarshani, None; Salma Ferdous, None; Shanu Markand, None; Kevin Donaldson, None; Jeffrey Boatright, None; J Nickerson, None
  • Footnotes
    Support  No
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 4550. doi:
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      Priyanka Priyadarshani, Salma Ferdous, Shanu Markand, Kevin Joseph Donaldson, Jeffrey H Boatright, J M Nickerson; Characterization of magnetofection transfection reagent, a novel transfection method for gene therapy in a mouse model. Invest. Ophthalmol. Vis. Sci. 2018;59(9):4550.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Improving the precise delivery of therapeutic nucleic acid at a specific target increases the activity at the desired area and improves precision of gene therapy, thus allowing us to fully utilize their potential in gene therapy. Previously we showed the magnetofection transfection system provides high throughput transfection efficiency both in vitro and in vivo model. The purpose of this study was to: 1) examine the efficiency of magnetofection reagent with and without presence of a magnet, 2) to assess the localization of GFP signal in the RPE cells, and 3) to examine the retinal function after transfection.

Methods : C57BL/6J mice at postnatal day 120-125 days were divided into two groups (n= 4/group). The right eye of each mouse was subretinally injected with gWiz_GFP plasmid mixed with PolyMag transfection reagent (Ozbiosciences) and the left eye was uninjected control. Concentration of each transfection reagents was used as per the manufacturer’s protocol. For the first group, the magnet was placed on the temporal area of mice for about 15-20 minutes after subretinal injection and for the second group, no magnet was placed after injection. Successful injection was validated using invivo fundus imaging i.e Micron IV system. Distribution and concentration of GFP in RPE flatmounts was observed using confocal microscopy. RPE flatmounts were analyzed for the localization and morphological changes after injection using Imaris Image Analysis Software. Post one month retinal functional was analyzed using electroretinogram (ERG).

Results : Bleb was evident after subretinal injection by fundus photograpgy and SDOCT. Confocal image analysis showed higher expression of GFP in the eye with magnet placed after injection. Imaris image analysis showed GFP with magnetic beads were mostly localized in the cytoplasm of the RPE cells. ERG analysis revealed normal retinal function in the mouse eye subretinally injected with magnetofection transfection reagent.

Conclusions : Our data suggest the presence of magnet impacted the efficiency of magnetofection transfection reagent and signal was localized in the specific targeted site without altering retinal functional.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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