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細胞穿透胜肽之分子運送 Molecular Delivery of Cell-Penetrating Peptides

細胞穿透胜肽之分子運送 Molecular Delivery of Cell-Penetrating Peptides. 李漢榮 國立東華大學 自然資源與環境學系. Speaker: Dr. Han-Jung Lee Department of Natural Resources and Environmental Studies National Dong Hwa University. b -Gal. Introduction. PTD. Extracellular. Intracellular. Plasma Membrane.

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細胞穿透胜肽之分子運送 Molecular Delivery of Cell-Penetrating Peptides

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  1. 細胞穿透胜肽之分子運送Molecular Delivery of Cell-Penetrating Peptides 李漢榮 國立東華大學自然資源與環境學系 Speaker: Dr. Han-Jung Lee Department of Natural Resources and Environmental Studies National Dong Hwa University

  2. b-Gal Introduction PTD Extracellular Intracellular Plasma Membrane Schwarze et al., Science 285: 1569-1572, 1999

  3. Cell-Penetrating Peptides (CPPs) Transactivator (Tat) protein of the human immunodeficiency virus type 1 (HIV-1) was able to enter into cells and activate HIV-LTR. Green and Loewenstein, Cell 55: 1179-1188, 1988 Frankel and Pabo, Cell 55: 1189-1193, 1988

  4. DNA RNA protein Tat O2 H2O Cell-Penetrating Peptides (CPPs) Extracellular Intracellular Plasma Membrane

  5. Cell-Penetrating Peptides (CPPs) Functional domains of HIV-1 Tat: Gaynor, Curr. Top. Microbiol. Immunol.193: 51-77, 1995 Vives et al., J. Biol. Chem. 272: 16010-16017, 1997 CxxCxCxxCCxxCxxC 1 11 22 37 72 2 47 57 86 cysteine-rich acidic basic core augmenting EPVDPRLEPW YGRKKRRQRRR Basic amino acid-rich domain

  6. Tat Cell-Penetrating Peptides (CPPs) CPPs, also called protein transduction domains (PTDs), are a group of short, highly basic peptides which can penetrate into cells either alone or associated with cargoes. YGRKKRRQRRR (CPP)

  7. Results Chang et al., Plant Cell Physiol. 46: 482-488, 2005

  8. GFP (200x) R9-GFP (200x) Tat-GFP (200x) R9-GFP (400x) Tat-GFP (400x) Covalent protein transduction into onion root tip cells

  9. R9-RFP RFP (tomato root cells) (onion epidermal cells) R9-RFP (onion epidermal cells) Covalent protein transduction in plant cells

  10. protein protein Covalent and Noncovalent Protein Transductions covalent protein transduction (CPT) noncovalent protein transduction (NPT) Plasma Membrane

  11. GFP R9/GFP RFP R9/RFP Noncovalent protein transduction in onion root cells Chang et al., New Phytol. 174: 46-56, 2007

  12. Cell-Penetrating Peptides (CPPs) CPPs are able to deliver proteins into cells in a covalent protein transduction (CPT) and noncovalent protein transduction (NPT).

  13. Cell-Penetrating Peptides (CPPs) • Synthetic nona-arginine (SR9) • N-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-C

  14. DNA SR9 (mung bean) (mung bean) SR9/DNA SR9/DNA (mung bean) (mung bean) DNA SR9/DNA (soybean) (soybean) Chen et al., FEBS Lett. 581: 1891-1897, 2007

  15. GC-EGFP cells dsRNA SR9/ dsRNA Wang et al., J. Biochem. Biophy. Methods 70: 579-586, 2007

  16. Cell-Penetrating Peptides (CPPs) CPPs can deliver proteins, DNAs and RNAs into cells in a noncovalent manner.

  17. GFP RFP PBS RFP R9-GFP R9-GFP R9-GFP/RFP R9-GFP/RFP Hu et al., Peptides 30: 1669-1678, 2009

  18. Cell-Penetrating Peptides (CPPs) CPPs are able to deliver proteins into cells in a covalent protein transduction (CPT), noncovalent protein transduction (NPT) and covalent and noncovalent protein transductions (CNPT).

  19. Quantum dots Lyso Tracker Hoechst (merge) Liu et al., Biomaterials 32: 3520-3537, 2011

  20. Summary We demonstrate that CPPs can effectively deliver proteins, DNAs, RNAs and nanoparticles into live cells. 17

  21. Summary However, the mechanism of protein transduction is unknown yet.

  22. CPP Translocation Mechanisms Cargo CPP

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