Enhanced delivery of protein fused to cell penetrating peptides to mammalian cells
1
조회
0
다운로드
4
인용
0
참고문헌
키워드
초록
Recent progress in cellular reprogramming technology and lineage-specific cell differentiation has provided great opportunities for translational research. Because virus-based gene delivery is not a practical reprogramming protocol, protein-based reprogramming has been receiving attention as a safe way to generate reprogrammed cells. However, the poor efficiency of the cellular uptake of reprogramming proteins is still a major obstacle. Here, we reported key factors which improve the cellular uptake of these proteins. Purified red fluorescent proteins fused with 9xLysine (dsRED-9K) as a cell penetrating peptide were efficiently delivered into the diverse primary cells. Protein delivery was improved by the addition of amodiaquine. Furthermore, purified dsRED-9K was able to penetrate all cell lineages derived from mouse embryonic stem cells efficiently. Our data may provide important insights into the design of protein-based reprogramming or differentiation protocols.
Abstract
Recent progress in cellular reprogramming technology and lineage-specific cell differentiation has provided great opportunities for translational research. Because virus-based gene delivery is not a practical reprogramming protocol, protein-based reprogramming has been receiving attention as a safe way to generate reprogrammed cells. However, the poor efficiency of the cellular uptake of reprogramming proteins is still a major obstacle. Here, we reported key factors which improve the cellular uptake of these proteins. Purified red fluorescent proteins fused with 9xLysine (dsRED-9K) as a cell penetrating peptide were efficiently delivered into the diverse primary cells. Protein delivery was improved by the addition of amodiaquine. Furthermore, purified dsRED-9K was able to penetrate all cell lineages derived from mouse embryonic stem cells efficiently. Our data may provide important insights into the design of protein-based reprogramming or differentiation protocols.
참고문헌 (23)
- Takahashi K, "Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors", Cell, 126, pp.663~676 (2006)
- Yamanaka S, "Strategies and new developments in the generation of patient-specific pluripotent stem cells", Cell Stem Cell, 1, pp.39~49 (2007)
- Okita K, "Generation of germline-competent induced pluripotent stem cells", Nature, 448, pp.313~317 (2007)
- Koren E, "Cell-penetrating peptides : breaking through to the other side", Trends Mol Med, 18, pp.385~393 (2012)
- El-Sayed A, "Delivery of macromolecules using arginine-rich cell-penetrating peptides : ways to overcome endosomal entrapment", AAPS J, 11, pp.13~22 (2009)
- Vives E, "A truncated HIV-1Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus", J Biol Chem, 272, pp.16010~16017 (1997)
- Herbig ME, "Membrane surfaceassociated helices promote lipid interactions and cellular uptake of human calcitonin-derived cell penetrating peptides", Biophys J, 89, pp.4056~4066 (2005)
- Wadia JS, "Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis", Nat Med, 10, pp.310~315 (2004)
- Nemes C, "Generation of mouse induced pluripotent stem cells by protein transduction", Tissue Eng Part C Methods, 20, pp.383~392 (2014)
- Kim D, "Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins", Cell Stem Cell, 4, pp.472~476 (2009)
- Li W, "Generation of rat and human induced pluripotent stem cells by combining genetic reprogramming and chemical inhibitors", Cell Stem Cell, 4, pp.16~19 (2009)
- Park H, "Increased genomic integrity of an improved protein-based mouse induced pluripotent stem cell method compared with current viral-induced strategies", Stem Cells Transl Med, 3, pp.599~609 (2014)
- Yang S, "Cellular uptake of self-assembled cationic peptide-DNA complexes : multifunctional role of the enhancer chloroquine", J Control Release, 135, pp.159~165 (2009)
- Shiraishi T, "Enhanced delivery of cell-penetrating peptide-peptide nucleic acid conjugates by endosomal disruption", Nat Protoc, 1, pp.633~636 (2006)
- Qiao S, "The antimalarial amodiaquine causes autophagic-lysosomal and proliferative blockade sensitizing human melanoma cells to starvation-and chemotherapy-induced cell death", Autophagy, 9, pp.2087~2102 (2013)
- Frankel AD, "Tat protein from human immunodeficiency virus forms a metal-linked dimer", Science, 240, pp.70~73 (1988)
- Rhee YH, "Protein-based human iPS cells efficiently generate functional dopamine neurons and can treat a rat model of Parkinson disease", J Clin Invest, 121, pp.2326~2335 (2011)
- Guidotti G, "Cell-Penetrating peptides : from basic research to clinics", Trends Pharmacol Sci, 38, pp.406~424 (2017)
- Kim CH, "Nuclear receptor Nurr1 agonists enhance its dual functions and improve behavioral deficits in an animal model of Parkinson's disease", Proc Natl Acad Sci U S A, 112, pp.8756~8761 (2015)
- Heidari R, "Amodiaquine-induced toxicity in isolated rat hepatocytes and the cytoprotective effects of taurine and", or N-acetyl cysteine, 2014, pp.9, 97~105
- Smidt MP, "How to make a mesodiencephalic dopaminergic neuron", Nat Rev Neurosci, 8, pp.21~32 (2007)
- Chung S, "The homeodomain transcription factor Pitx3 facilitates differentiation of mouse embryonic stem cells into AHD2-expressing dopaminergic neurons", Mol Cell Neurosci, 28, pp.241~252 (2005)
- Andersson E, "Identification of intrinsic determinants of midbrain dopamine neurons", Cell, 124, pp.393~405 (2006)