title: Delivery of cell-penetrating peptide-peptide nucleic acid conjugates by assembly on an oligonucleotide scaffold.
abstract: Delivery to intracellular target sites is still one of the main obstacles in the development of peptide nucleic acids (PNAs) as antisense-antigene therapeutics. Here, we designed a self-assembled oligonucleotide scaffold that included a central complementary region for self-assembly and lateral regions complementing the PNAs. Assembly of cell-penetrating peptide (CPP)-PNAs on the scaffold significantly promoted endocytosis of PNAs by at least 10-fold in cell cultures, particularly for scaffolds in which the central complementary region was assembled by poly(guanine) and poly(cytosine). The antisense activity of CPP-PNAs increased by assembly on the scaffold and was further enhanced after co-assembly with endosomolytic peptide (EP)-PNA. This synergistic effect was also observed following the assembly of antigene CPP-PNAs\EP-PNAs on the scaffold. However, antigene activity was only observed by targeting episomal viral DNA or transfected plasmids, but not the chromosome in the cell cultures. In conclusion, assembly on oligonucleotide scaffolds significantly enhanced the antisense-antigene activity of PNAs by promoting endocytosis and endosomal escape. This oligonucleotide scaffold provided a simple strategy for assembly of multiple functional peptide-PNA conjugates, expanding the applications of PNAs and demonstrating the potential of PNAs as antiviral therapeutics.
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases; Beijing 100044, P.R. China.
authors.forename: Xing-Liang
authors.initials: XL
authors.lastname: Zhao
authors.affiliation: Zhejiang Provincial Top Key Discipline in Surgery, Wenzhou Key Laboratory of Surgery; Department of Surgery, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou, 325200, P.R. China.
authors.forename: Bi-Cheng
authors.initials: BC
authors.lastname: Chen
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases; Beijing 100044, P.R. China.
authors.forename: Jin-Chao
authors.initials: JC
authors.lastname: Han
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases; Beijing 100044, P.R. China.
authors.forename: Lai
authors.initials: L
authors.lastname: Wei
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases; Beijing 100044, P.R. China.
authors.forename: Xiao-Ben
authors.initials: XB
authors.lastname: Pan
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D057846:Cell-Penetrating Peptides / Q000138:chemical synthesis / Q000235:genetics* / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000235:genetics* / Q000378:metabolism; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011066:Poly C / Q000737:chemistry / Q000378:metabolism; D011068:Poly G / Q000737:chemistry / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; D057846:Cell-Penetrating Peptides; D004279:DNA, Viral; D016376:Oligonucleotides, Antisense; D020135:Peptide Nucleic Acids; D011068:Poly G; D011066:Poly C