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NOVEL CARRIER MOLECULE AND METHOD BOTH FOR DELIVERING RNA INTO CYTOPLASM USING NEAR-INFRARED LIGHT コモンズ

外国特許コード F130007322
整理番号 S2011-0272-C0
掲載日 2013年4月26日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2011JP077075
国際公開番号 WO 2012127739
国際出願日 平成23年11月24日(2011.11.24)
国際公開日 平成24年9月27日(2012.9.27)
優先権データ
  • 特願2011-063953 (2011.3.23) JP
発明の名称 (英語) NOVEL CARRIER MOLECULE AND METHOD BOTH FOR DELIVERING RNA INTO CYTOPLASM USING NEAR-INFRARED LIGHT コモンズ
発明の概要(英語) The purpose of the present invention is to provide a means for introducing RNA into a cell utilizing near-infrared light that has good biological tissue permeability (preferably having a wavelength of about 700-1000 nm) without causing the problem of cytotoxicity or heat generation. A carrier molecule according to the present invention is characterized by having a structure comprising: a carrier protein which comprises a cell-membrane-permeable peptide (CPP) and an RNA-binding protein (RBP); and a photosensitizer (PST) which is connected to the N-terminal side or the C-terminal side of the carrier protein and can act upon the irradiation with light having a wavelength falling within a near infrared region. A (carrier molecule)/RNA complex composed of the carrier molecule and RNA bound to the RBP in the carrier molecule is formed, the complex is brought into contact with a cell to localize the complex in an endosome in the cell, and subsequently the cell is irradiated with near infrared light with which the PST in the (carrier molecule)/RNA complex can act. In this manner, the (carrier molecule)/RNA complex can be dispersed in a cytoplasm, whereby the function of the introduced RNA can be exerted.
従来技術、競合技術の概要(英語) BACKGROUND ART
Is RNAi, siRNA or shRNA depending on the nucleotide sequence of a particular gene expression is repressed phenomenon, disease-causing gene or suppression of expression of the viral genes can be used in applications such as disease treatment based on very recent years has attracted attention. In recent years, by generating ions and emitting, relieve stress or discomfort, virus and fungi, allergens cause allergic diseases or rendered harmless by the ion generator can be employed in various applications and is, for example, an unspecified number of persons used by the ion generating device placed in the elevator has been a growing demand for.
Is Patent Document 1, membrane permeable carrier peptide RNA binding protein and a fusion protein, the recognition sequence for the RNA-binding protein having a sequence and any bound RNA, the conjugate of any of the cells by mixing, the RNA introduced into the cell and method described.
Non-patent document 1 and 2 is, the carrier peptide RNA binding protein and membrane permeability in addition to the wavelength in the visible region excited with light of a fluorescent dye (for example Alexa Fluor 546, 530-550 nm excitation wavelength) to a fusion protein comprising siRNA bound to the complex is described. Then, contacting a cell with this conjugate, the conjugate incorporated into cells by endocytosis after localization to the endosome, by irradiating the excitation light, the fluorescent dye acts as a photosensitizer, is released into the cytoplasm from the endosome siRNA, a predetermined RNAi effect can be achieved (see Fig. 1) described. In addition, non-patent document 1 and 2 is, near-infrared region of wavelength excited with light of a fluorescent pigment may also be used to obtain the above-mentioned effect can be achieved is, not describe or suggest.
On the other hand, Patent Document 2 is, as a carrier of a functional molecule gold nano-rods, gold nano-rod complex aggregates DNA and added to the cultured cell, a near-infrared region (wavelength 1064 nm) by the pulse laser is irradiated, the gold nanorods to eliminate DNA, effective expression of the DNA of the gene from the described method (see Examples). However, the photothermal conversion function and has gold nano-rods may be, gold nano-rods are integrated on a photo thermal treatment of the surrounding tissue to the target site (tumor cells kill heat generated) is also used for the material (see for example Patent Document 3), as such is not intended for use in the cytotoxicity or concern for safety.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • NATIONAL UNIVERSITY CORPORATION OKAYAMA UNIVERSITY
  • 発明者(英語)
  • OHTSUKI, Takashi
  • ISHIODORI, Yuka
国際特許分類(IPC)
指定国 National States: AE AG AL AM AO AT AU AZ BA BB BG BH BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IS JP KE KG KM KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PE PG PH PL PT QA RO RS RU RW SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ MD RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

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