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Designing of DLinDMA‐Anchored Liposomal Carrier for Gene Therapy of Rare Genetic Lafora Disease
Author(s) -
Vemana Hari Priya,
Saraswat Aishwarya,
Bhutkar Shraddha,
Patel Ketan,
Dukhande Vikas
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.04021
Subject(s) - transfection , cationic liposome , microbiology and biotechnology , chemistry , liposome , biochemistry , biology , gene
Lafora disease (LD) is a fatal, autosomal recessive, progressive myoclonic epilepsy. The disease occurs due to loss of function in phosphatase laforin or E3 ligase malin. There is only palliative treatment available for LD. The promise of gene therapeutics to treat such lethal conditions led us to develop a novel type of cationic liposomes as non‐viral vectors to encapsulate plasmid DNA containing EPM2A gene encoding laforin. Cationic liposomes containing DLinDMA were formulated and characterized for their particle size and zeta potential. DLinDMA liposomes were tested for their cytotoxicity in HEK 293 and neuroblastoma cells. Gel retardation assay was performed to analyze the electrophoretic mobility of the DNA containing lipoplexes. Fluorescence microscopy studies showed that the transfection efficiency of plasmids containing EGFP or laforin encapsulated liposomes was comparable to commonly used transfection reagents such as Transit LT‐1 and PEI‐Max. The expression of laforin was confirmed using Western blotting. Furthermore, malachite green based glucan phosphatase assay was performed to assess the biological activity of laforin. Further in vivo studies will be performed to monitor pharmacokinetics and biological activity. Our novel study highlights the importance of DLinDMA cationic lipoplexes as a promising non‐viral vector to efficiently deliver plasmid DNA for the treatment of fatal rare genetic diseases such as LD.