Smooth muscle cell targeting gene delivery using artery wall binding peptide linked high mobility group box 1 A

Abstract

High mobility group box 1(HMGB1) are able to condense with plasmid DNA. Previously we generated a truncated HMGB1 derivative which lacks the acidic C-tail, reported to decrease the affinity of HMG proteins for DNA, and HMG box B. HIV-TAT protein was conjugated to truncated formation of HMGB1(HMGB1A) to enhance transfection efficiency. Apolipoprotein B-100 contains artery wall binding domain. In this study, artery wall binding peptide (ABP) was conjugated to TAT-HMGB1A for artery wall targeting and it enhance gene expression. Recombinant his tag fusion protein, TAT-HMG1A-ABP were cloned and purified. The complexes formation between TAT-HMGB1A-ABP and plasmid DNA was confirmed by gel retardation assay. pCMV-Luc and peptide complexes were added to A7R5 smooth muscle. The highest transfection efficiency of TAT-HMGB1A-ABP was at a 20/1 weight ratio. TAT-HMGB1A-ABP/pDNA complex had higher efficiency than PLL. In competition assay with free ABP, the transfection of TAT-HMGB1A-ABP was decreased by the addition of free ABP at a concentration of 300 ug/ml. MTT assay was performed to investigate whether protein carrier is non-toxic. The Recombinant protein for artery wall targeting has considerable potential to facilitate DNA delivery to smooth muscle cells.