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Recombinant derivatives of the human high‐mobility group protein HMGB2 mediate efficient nonviral gene delivery
Author(s) -
Sloots Arjen,
Wels Winfried S.
Publication year - 2005
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2005.04834.x
Subject(s) - transfection , fusion protein , biology , recombinant dna , gene delivery , microbiology and biotechnology , gene , akt1s1 , plasmid , fusion gene , gatad2b , genetic enhancement , importin , transduction (biophysics) , hspa9 , hspa2 , biochemistry , nuclear transport , peptide sequence , cell nucleus
Certain natural peptides and proteins of mammalian origin are able to bind and condense plasmid DNA, a prerequisite for the formation of transfection‐competent complexes that facilitate nonviral gene delivery. Here we have generated recombinant derivatives of the human high‐mobility group (HMG) protein HMGB2 and investigated their potential as novel protein‐based transfection reagents. A truncated form of HMGB2 encompassing amino acids 1–186 of the molecule was expressed in Escherichia coli at high yield. This HMGB2 186 protein purified from bacterial lysates was able to condense plasmid DNA in a concentration‐dependent manner, and mediated gene delivery into different established tumor cell lines more efficiently than poly( l ‐lysine). By attaching, via gene fusion, additional functional domains such as the HIV‐1 TAT protein transduction domain (TAT PTD ‐HMGB2 186 ), the nuclear localization sequence of the simian virus 40 (SV40) large T‐antigen (SV40 NLS ‐HMGB2 186 ), or the importin‐β‐binding domain (IBB) of human importin‐α (IBB‐HMGB2 186 ), chimeric fusion proteins were produced which displayed markedly improved transfection efficiency. Addition of chloroquine strongly enhanced gene transfer by all four HMGB2 186 derivatives studied, indicating cellular uptake of protein–DNA complexes via endocytosis. The IBB‐HMGB2 186 molecule in the presence of the endosomolytic reagent was the most effective. Our results show that recombinant derivatives of human HMGB2 facilitate efficient nonviral gene delivery and may become useful reagents for applications in gene therapy.