Gene transfer into human hepatoma cells by receptor-associated protein/polylysine conjugates
DC Field | Value | Language |
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dc.contributor.author | Kim, TG | - |
dc.contributor.author | Kang, SY | - |
dc.contributor.author | Kang, JH | - |
dc.contributor.author | Cho, MY | - |
dc.contributor.author | Kim, JI | - |
dc.contributor.author | Kim, SH | - |
dc.contributor.author | Kim, JS | - |
dc.date.available | 2021-02-22T16:03:10Z | - |
dc.date.issued | 2004-03 | - |
dc.identifier.issn | 1043-1802 | - |
dc.identifier.issn | 1520-4812 | - |
dc.identifier.uri | https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/15903 | - |
dc.description.abstract | Receptor-associated protein (RAP) is a ligand for all members of low-density lipoprotein (LDL) receptor families. RAP is internalized into cells via receptor-mediated endocytic trafficking, making it an attractive mechanism for efficient gene delivery. In this study, we have developed a gene delivery system using RAP as a targeting ligand. A RAP cDNA lacking a C-terminal heparin-binding domain was amplified by polymerase chain reaction (PCR) from a human liver cDNA library and was reamplified by using a primer containing a cysteine codon at its carboxyl end to facilitate its conjugation to polylysine (polyK). RAP was purified using a bacterial expression system and coupled to poly-D-lysine (PDL) or poly-L-lysine (PLL) of average MW 50 kDa via the heterobifunctional cross-linker SPDP. Using fluorescence-labeled RAP ligand, cellular uptake of the transfection complexes into HepG2 cells was shown to be highly efficient and more specific to PDL-conjugated RAP compared with PLL-conjugated one. Plasmid DNA containing a luciferase reporter gene was condensed with either RAP-PDL or RAP-PLL. In vitro transfection into HepG2 cells with RAP-PDL conjugate resulted in significantly higher luciferase expression levels in comparison to either nonconjugated PDL, or RAP-PLL, or LipofecAMINE/DNA complexes in the presence of 10% fetal bovine serum. Luciferase expression was inhibited by the addition of excess RAP. Treatment of the cells with Lovastatin, which inhibits HMG-Co reductase and increases expression of LDL receptor, stimulates luciferase expression, suggesting that the gene delivery is specifically mediated by LDL receptor. Thus, RAP-PDL conjugates have the potential to be used as a new nonviral gene delivery vector. | - |
dc.format.extent | 7 | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | AMER CHEMICAL SOC | - |
dc.title | Gene transfer into human hepatoma cells by receptor-associated protein/polylysine conjugates | - |
dc.type | Article | - |
dc.publisher.location | 미국 | - |
dc.identifier.doi | 10.1021/bc0340262 | - |
dc.identifier.scopusid | 2-s2.0-1642312927 | - |
dc.identifier.wosid | 000220284100012 | - |
dc.identifier.bibliographicCitation | BIOCONJUGATE CHEMISTRY, v.15, no.2, pp 326 - 332 | - |
dc.citation.title | BIOCONJUGATE CHEMISTRY | - |
dc.citation.volume | 15 | - |
dc.citation.number | 2 | - |
dc.citation.startPage | 326 | - |
dc.citation.endPage | 332 | - |
dc.type.docType | Article | - |
dc.description.isOpenAccess | N | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Chemistry | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Multidisciplinary | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Organic | - |
dc.subject.keywordPlus | LOW-DENSITY-LIPOPROTEIN | - |
dc.subject.keywordPlus | MEDIATED ENDOCYTOSIS | - |
dc.subject.keywordPlus | DELIVERY SYSTEM | - |
dc.subject.keywordPlus | CARCINOMA-CELLS | - |
dc.subject.keywordPlus | DNA COMPLEXES | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | PROTEIN | - |
dc.subject.keywordPlus | CANCER | - |
dc.subject.keywordPlus | LIGAND | - |
dc.subject.keywordPlus | TRANSFORMATION | - |
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