• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

神経幹細胞への染色体部位特異的遺伝子組込み法の開発

Research Project

Project/Area Number 15659343
Research Category

Grant-in-Aid for Exploratory Research

Allocation TypeSingle-year Grants
Research Field Cerebral neurosurgery
Research InstitutionJichi Medical University

Principal Investigator

岡田 尚已  自治医科大学, 医学部, 講師 (00326828)

Co-Investigator(Kenkyū-buntansha) 小澤 敬也 (小沢 敬也)  自治医科大学, 医学部, 教授 (30137707)
卜部 匡司  自治医科大学, 医学部, 講師 (40213516)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2004: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
Keywordsアデノ随伴ウイルス / 染色体部位特異的遺伝子組込み / Rep / AAVS1 / 幹細胞 / アデノウイルスベクター / CAR / c-Kit / 染色体遺伝子組込み / アデノウイルス
Research Abstract

AAVの二つのコンポーネント(ITR配列とRep蛋白質)を利用し、第19番染色体長腕AAVS1領域に遺伝子を組み込ませる方法の応用開発を推進した。これまでにITR配列連結マーカー発現ユニットを構築し、切り出されたstuffer配列中のITRにRepが結合することによって、第19番染色体AAVS1領域に遺伝子組込みが生ずることを検討した。HeLa細胞を用いて、Southern blot解析によるAAVS1領域の遺伝子再構成とマーカー遺伝子のAAVS1領域特異的遺伝子組込み様式を確認した。
さらに、ITR配列連結マーカー発現ユニットとRep発現誘導カセットを有するアデノウイルスベクターの応用を念頭に、アデノウイルスベクターの幹細胞への吸着を仲介するアダプター分子を開発した。アデノウイルスベクターに親和性のあるCARと幹細胞に発現するc-Kitのリガンド分子である幹細胞因子(SCF)を連結した、可溶性融合蛋白質CAR-SCFを構築し、その効果を検討した。CAR陰性c-Kit陽性の細胞株M07eへのeGFP発現Adベクター遺伝子導入(MOI 50)において、CAR-SCFを培養液に添加することでeGFP陽性率は0.5%から78%に向上した。ヒト臍帯血CD34^+細胞を用いた検討でも、9.8±3.7%から17.5±2.5%に改善された。CAR-SCFはc-Kit陽性細胞へのアデノウイルスベクター遺伝子導入効率を向上させ、これを用いた幹細胞へのAAVS1領域特異的遺伝子組込みへの応用が期待される。

Report

(2 results)
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (13 results)

All 2005 2004 2003 Other

All Journal Article (6 results) Patent(Industrial Property Rights) (1 results) Publications (6 results)

  • [Journal Article] Efficient and stable Sendai virus-mediated gene transfer into primate embryonic stem cells with pluripotency preserved.2005

    • Author(s)
      Sasaki, K., et al.
    • Journal Title

      Gene Ther. 12・3

      Pages: 203-210

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Separate Control of Rep and Cap Expression Using Mutant and Wild-Type LoxP Sequences and Improved Packaging System for Adeno-Associated Virus Vector Production.2004

    • Author(s)
      Mizukami, H., et al.
    • Journal Title

      Mol Biotechnol. 27・1

      Pages: 7-14

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Expansion of genetically corrected neutrophils in chronic granulomatous disease mice by cotransferring a therapeutic gene and a selective amplifier gene.2004

    • Author(s)
      Hara, T., et al.
    • Journal Title

      Gene Ther 11・18

      Pages: 1370-1377

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Establishment of Cre/LoxP recombination system in transgenic rats.2004

    • Author(s)
      Sato, Y., et al.
    • Journal Title

      Biochem Biophys Res Commun. 319・4

      Pages: 1197-1202

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A soluble CAR-SCF fusion protein improves adenoviral vector-mediated gene transfer to c-Kit-positive hematopoietic cells.2003

    • Author(s)
      Itoh, A., et al.
    • Journal Title

      J.Gene Med. 5・11

      Pages: 929-940

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Development of new inbred transgenic strains of rats with LacZ or GFP.

    • Author(s)
      Inoue, H., et al.
    • Journal Title

      Biochem Biophys Res Commun. (in press)

    • Related Report
      2004 Annual Research Report
  • [Patent(Industrial Property Rights)] Agents for adsorption and ridging for adenovirus2003

    • Inventor(s)
      Itoh A., et al.
    • Industrial Property Rights Holder
      旭化成
    • Filing Date
      2003-05-15
    • Related Report
      2004 Annual Research Report
  • [Publications] Okada T, et al.: "In situ generation of pseudotyped retroviral progeny by adenovirus-mediated transduction of tumor cells enhances the killing effect of HSV- tk suicide gene therapy in vitro and in vivo"J.Gene Med.. (in press).

    • Related Report
      2003 Annual Research Report
  • [Publications] Itoh A, et al.: "A soluble CAR-SCF fusion protein improves adenoviral vector-mediated gene transfer to c-Kit-positive hematopoietic cells"J.Gene Med.. 5・11. 929-940 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nomoto T, et al.: "Distinct patterns of gene transfer to gerbil hippocampus with recombinant adeno-associated virus type 2 and 5"Neuroscience Letters. 340. 153-157 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Lu Y Y, et al.: "Intramuscular injection of AAV-GDNF results in sustained expression of transgenic GDNF, and its delivery to spinal motoneurons by retrograde transport"Neurosci Res.. 45・1. 33-40 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ideno J, et al.: "Persistent phenotypic correction of central diabetes insipidus using adeno-associated virus vector-expressing arginine-vasopressin in brattleboro rats"Mol.Ther.. 8・6. 895-902 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kume A, et al.: "In vivo expansion of transduced murine hematopoietic cells with a selective amplifier gene"J.Gene Med.. 5・3. 175-181 (2003)

    • Related Report
      2003 Annual Research Report

URL: 

Published: 2003-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi