An archaeal histone-like protein mediates efficient p53 gene transfer and facilitates its anti-cancer effect in vitro and in vivo

Abstract

The improvement of the transfection efficiency of the non-viral-based gene delivery systems is a key issue for the application in gene therapy. We have previously described an archaeal histone-like protein-based (HPhA) gene delivery system and showed that HPhA formed stable non-covalent complexes with nucleic acids and improved their delivery by using b-galactosidase as a reporter gene. In this study, the wild-type p53 gene was transfected into the cancer cells using the HPhA as a vector, and the expression level and the activity of p53 gene were evaluated both in vitro and in vivo. Gene expression was determined by real-time reverse transcriptase-PCR and western blotting analysis. The cellular growth inhibition and apoptosis of HPhA-mediated p53 transfection were assessed by XTT (sodium 3 0 -[1-(phenylaminocarbonyl)-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzene sulfonic acid hydrate) assay and annexin V-FITC (fluorescein isothiocyanate) staining, respectively. Further more, transfection of HPhA/p53 into CNE (nasopharyngeal carcinoma cell line)-xenografted nude mice was performed and tumor growth was measured. The present study demonstrates that HPhA enhances the efficiency of p53 gene transfer and antitumor activity compared with the widely used Lipofectamine. These results demonstrate that HPhA enhances the in vitro and in vivo efficiency of p53 gene transfer and suggest that it may be served as a promising tool for gene delivery and gene therapy.

Genipin-Cross-Linked Thermophilic Histone-Polyethylenimine as a Hybrid Gene Carrier

“…In addition, histone-mediated transmembrane was performed in a direct translocation manner (not by a typical endocytosis), which would facilitate the endosomal escape of carrier/DNA nanocomplexes. , More importantly, synergistic effects have been observed between histones and polymeric gene carriers, such as PEI and its derivative poly­( l -glutamic acid)- g -PEI. − However, most of these histones were obtained from calf thymus or chick erythrocyte, and immune response was difficult to be avoided due to their intrinsic characteristic of high molecular weight. Compared with histones of these originations, thermophilic histones from thermophiles are potential to be efficient gene carriers with superior characteristics: − (1) they possess the basic fold of eukaryotic histones, and thus hold favorable DNA binding and package ability; (2) their molecular weights are much lower (<10 kDa), which will be favorable for decreasing the immunogenicity and cytotoxicity; and (3) they exhibit high stability against thermal, organic solvents, and denaturants, which will be beneficial for further modification or conjugating with other carriers, especially under harsh reaction conditions. Nevertheless, their transfection efficiencies are still lower than polymeric gene carriers, and thus, it is necessary to construct a thermophilic histone–polymer hybrid gene carrier for improving the transfection efficiency and combining their individual advantages.…”

Inhibition of HCV 5′-NTR and Core Expression by a Small Hairpin RNA Delivered by a Histone Gene Carrier, HPhA

“…The core of the nuclear body is histone, wound tightly by DNA. However, the tight binding can loosen during DNA binding to other proteins such as polymerase 25,29,30.…”

Diiron Dithiolate Complex Induced Helical Structure of Histone and Application in Photochemical Hydrogen Generation

“…Histone H1 consists of three distinct domains: a short N-terminal domain (NTD) (20–35 amino acids), a stably folded central globular domain (∼80 amino acids), and a long C-terminal domain (CTD) (∼100 amino acids). , The globular domain is extremely conserved, while the terminal domains, in particular, the CTD, are very rich in lysine (∼40%), fully mobile and intrinsically unstructured in aqueous solution. − As the CTD has a high helix potential, the neutralization of lysine positive charges by chemical modifications can induce the folding of the CTD with proportions of secondary structure motifs. ,, Herein, we selected commercially available calf thymus histone H1 as the carrier of [FeFe] catalyst because of its exceptional stability of peptide backbones, plenty of lysine residues for immobilization, and most importantly the self-assembly property that allows for extensive study on the protein conformational changes and aggregates. − As shown in Scheme , the active ester [Fe 2 (lip-NHS)­(CO) 6 ] (Fe 2 S 2 –NHS) was conjugated to the lysine residues of disordered histone proteins to form histone- g -Fe 2 S 2 (H–Fe) nanoparticles after a reassembly process. Different from apoferritin encapsulating [FeFe] catalyst via coordination interaction, the linkage of histone with [FeFe] catalyst is dominated by covalent amide bonding to ensure no leakage of catalyst in broad pH ranges and induce a conversion of histone proteins from disordered structure to helical structure.…”