Vault nanoparticles containing an adenovirus-derived membrane lytic protein facilitate toxin and gene transfer.
Nonviral methods of gene delivery possess several advantages over that of viral-based vectors, including having increased safety. However, the ability to achieve effective transport of therapeutic molecules across host cell membranes via nonviral methods remains a significant goal. Cell-derived nanoparticles known as vaults have been proposed as novel candidate transfer vehicles for various foreign molecules. Recombinant vault particles enter cells via macropinocytosis or phagocytosis but lack demonstrable membrane penetrating activity. To explore the feasibility of improving vault penetration into target cells, we incorporated the membrane lytic domain of adenovirus protein VI (pVI) into the interior of recombinant vault particles via fusion to the vault poly(ADP-ribose) polymerase (VPARP) interaction domain. The membrane lytic activity of the pVI domain was retained upon incorporation into vault particles. Moreover, internalization of vault-pVI complexes into murine macrophages promoted co-delivery of a soluble ribotoxin or a cDNA plasmid encoding GFP. These findings indicate that vault particles can be modified to enhance cell transfer of selected biomolecules.
Lai
Cheng-Yu
CY
Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California 92037, USA.
Wiethoff
Chris M
CM
Kickhoefer
Valerie A
VA
Rome
Leonard H
LH
Nemerow
Glen R
GR
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D018014:Gene Transfer Techniques; D006801:Humans; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry*; D010446:Peptide Fragments / Q000235:genetics; D011065:Poly(ADP-ribose) Polymerases / Q000737:chemistry; D017434:Protein Structure, Tertiary; D020394:Vault Ribonucleoprotein Particles / Q000737:chemistry; D014764:Viral Proteins / Q000235:genetics
D001427:Bacterial Toxins; D010446:Peptide Fragments; D020394:Vault Ribonucleoprotein Particles; D014764:Viral Proteins; C105093:glycyl-valyl-glutaminyl-seryl-leucyl-lysyl-arginyl-arginyl-arginyl-cysteinyl-phenylalanine; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases; C449359:vault poly(ADP-ribose) polymerase