Poly(β-amino ester)-nanoparticle mediated transfection of retinal pigment epithelial cells in vitro and in vivo

Joel C. Sunshine, Sarah B. Sunshine, Imran Bhutto, James Handa, Jordan Green

Research output: Contribution to journalArticle

Abstract

A variety of genetic diseases in the retina, including retinitis pigmentosa and leber congenital amaurosis, might be excellent targets for gene delivery as treatment. A major challenge in non-viral gene delivery remains finding a safe and effective delivery system. Poly(beta-amino ester)s (PBAEs) have shown great potential as gene delivery reagents because they are easily synthesized and they transfect a wide variety of cell types with high efficacy in vitro. We synthesized a combinatorial library of PBAEs and evaluated them for transfection efficacy and toxicity in retinal pigment epithelial (ARPE-19) cells to identify lead polymer structures and transfection formulations. Our optimal polymer (B5-S5-E7 at 60 w/w polymer:DNA ratio) transfected ARPE-19 cells with 44±5% transfection efficacy, significantly higher than with optimized formulations of leading commercially available reagents Lipofectamine 2000 (26±7%) and X-tremeGENE HP DNA (22±6%); (p0.05 for both). Our optimal polymer was also significantly better than a gold standard polymeric transfection reagent, branched 25 kDa polyethyleneimine (PEI), which achieved only 8±1% transfection efficacy with 25±6% cytotoxicity. Subretinal injections using lyophilized GFP-PBAE nanoparticles resulted in 1.1±1×103-fold and 1.5±0.7×103-fold increased GFP expression in the retinal pigment epithelium (RPE)/choroid and neural retina respectively, compared to injection of DNA alone (p = 0.003 for RPE/choroid, p

Original languageEnglish (US)
Article numbere37543
JournalPLoS One
Volume7
Issue number5
DOIs
StatePublished - May 21 2012

Fingerprint

Retinal Pigments
nanoparticles
transfection
Nanoparticles
Transfection
Polymers
Esters
epithelial cells
Epithelial Cells
esters
pigments
polymers
gene transfer
Genes
Choroid
Retinal Pigment Epithelium
DNA
retina
Retina
Polyethyleneimine

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Poly(β-amino ester)-nanoparticle mediated transfection of retinal pigment epithelial cells in vitro and in vivo. / Sunshine, Joel C.; Sunshine, Sarah B.; Bhutto, Imran; Handa, James; Green, Jordan.

In: PLoS One, Vol. 7, No. 5, e37543, 21.05.2012.

Research output: Contribution to journalArticle

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