Evaluation of the use of amphipathic peptide-based protein carrier for in vitro cancer research.
Intracellular delivery of proteins offers an alternative to genetic modification or siRNA transfection for functional studies of proteins in live cells, especially for studies in cancer cells for therapeutics development. However, lack of efficient and biocompatible delivery system has limited the use of protein for in vitro cancer research. In this study, we design and evaluate an amphipathic peptide RV24, composing of a hydrophobic domain for protein binding, a flexible linker, and a hydrophilic domain to facilitate cell penetration. When using β-galactosidase as a cargo protein for comparison with commercially available peptide- and lipid-based carriers, RV24 peptide provides up to 5-fold increase in quantity delivered into 3 different cancer cell lines. Green fluorescent protein could also be delivered rapidly within 4h and transduced up to 83% of tested cancer cell lines. Although having a cell penetrating domain, RV24 peptide did not compromise cell viability, morphology and granularity significantly. These findings suggest the feasibility of using biocompatible amphipathic peptide to efficiently deliver protein-based molecules intracellularly for in vitro cancer research.
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, 138669 Singapore, Singapore. sllo@ibn.a-star.edu.sg
Seong Loong
SL
Lo
Shu
S
Wang
D004337:Drug Carriers / Q000737:chemistry / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D009369:Neoplasms / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D021381:Protein Transport; D011506:Proteins / Q000378:metabolism*; D001616:beta-Galactosidase / Q000378:metabolism
D004337:Drug Carriers; D010455:Peptides; D011506:Proteins; C571743:RV24 peptide; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase