Gene delivery using a derivative of the protein transduction domain peptide, K-Antp.
Due to their intracellular permeability, protein transduction domains (PTDs) have been widely used to deliver proteins and peptides to mammalian cells. However, their performance in gene delivery has been relatively poor. To improve the efficiency of PTD-mediated gene delivery, we synthesized a new peptide, KALA-Antp (K-Antp), which contains the sequences for PTD of the third alpha-helix of Antennapedia (Antp) homeodomain and the fusogenic peptide KALA. In this configuration, Antp is designed to provide the cell permeation capacity and nuclear localization signal, while the KALA moiety to promote cellular entry of the peptide-DNA complex. An optimal K-Antp/DNA formula was nearly 400-600 fold more efficient than Antp or poly-lysine-Antp (L-Antp) in gene delivery, and comparable or superior to a commercial liposome. The K-Antp-mediated plasmid DNA transfection not only exhibited temperature sensitivity, reflecting the involvement of an endocytosis-mediated gene transfer mechanism similar to other known PTDs, but also temperature insensitivity, suggesting the role of an energy-independent mechanism. Incorporation of an endosomolytic polymer polyethylenimine (PEI) into the system or treatment with chloroquine further increased the efficiency of K-Antp-mediated gene delivery. These results demonstrate the potential of the combinatorial use of KALA, Antp and PEI in the development of efficient PTD-derived gene carriers.
Min
Sang-Hyun
SH
Medical Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology, 111 Gwahangno, Yuseong-gu, Daejeon 305-806, Republic of Korea.
Kim
Dong Min
DM
Kim
Mi Na
MN
Ge
Jiang
J
Lee
Dong Chul
DC
Park
In Young
IY
Park
Kyung Chan
KC
Hwang
Ji-Sook
JS
Cho
Cheong-Weon
CW
Yeom
Young Il
YI
D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics; D011992:Endosomes / Q000378:metabolism; D018014:Gene Transfer Techniques; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011108:Polymers / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013696:Temperature; D014162:Transfection
C548457:KALA-Antp peptide; D010455:Peptides; D011108:Polymers; D004247:DNA