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High efficient transduction of renal tubular cells by Tyrosine‐mutant AAV2 sextuple vector
Author(s) -
Qi Yanfei,
Li QiuHong,
Shenoy Vinayak,
Jun JooYun,
Verma Amrisha,
Katovich Michael J,
Raizada Mohan K
Publication year - 2011
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.25.1_supplement.lb620
Subject(s) - transduction (biophysics) , green fluorescent protein , mutant , biology , microbiology and biotechnology , reporter gene , genetic enhancement , kidney , viral vector , gene , capsid , virology , virus , gene expression , genetics , biochemistry , recombinant dna
Gene therapy has distinct potential to treat kidney diseases; however efficient transduction of significant number of cells in the renal tissue by viral vectors has been difficult to accomplish. Previous studies have indicated that different serotypes of adeno‐associated virus (AAV) can transduce renal cells with variable and suboptimal efficiency. The availability of new and innovative mutants of AAV has led us to compare their efficacy in transducing rat kidney. Five types of AAV (AAV2 muttriple, AAV2 sextuple, AAV8 mut44, AAV8 mut733, and AAV9 mut446) carrying an enhanced green fluorescence protein (EGFP) reporter gene under the control of small chicken β‐actin (AAV‐CBA‐EGFP) were compared, which contain single or multiple point mutations in surface‐exposed capsid tyrosine residues. 1.5x10 11 vector genome (vg) of AAV in 20μl was directly injected into the renal cortex at three different sites using a pressure microinjection technique. Three weeks after injection, the kidneys were collected and evaluated for EGFP expression. Among the various mutated AAV serotypes studied, only AAV2 sextuple showed a robust EGFP expression in significant renal tissue. About 45% of the renal tubular epithelial cells expressed EGFP. Our study shows that mutated AAV2 sextuple is an efficient gene transfer vector for renal tissue and may hold future therapeutic applications to treat kidney diseases.