DNA delviery to artery wall cells with a targeting ligand linked recombinant high mobility group box-1 peptide

Abstract

HMGB1(high mobility group box 1) are able to bind 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 many receptor binding domain, including artery wall binding domain. In this study, artery wall binding peptide (ABP) was oncjugated to TAT-HMGB1A for artery wall targeting and it enhance gene expression. Recombinant His tag fusion protein, TAT-HMGB1A-ABP were cloned and purified. The complexes formation between TAT-HMGB1A-ABP and plasmid DNA was cofirmed by gel retardation assay. pCMV-Luc and peptide complexes were added to A7R5 smooth muslce. 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. MTT assay was performed to investigated whether protein carrier is non-toxic. The recombinant protein for artery wall targeting has considerable potential to facilitate DNA delivery to smooth muscle cells.