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著作: Kyoung Ho Lee/[鬼塚 正義]/Honda Kohsuke/Ohtake Hisao/[大政 健史]/Rapid construction of transgene-amplified CHO cell lines by cell cycle checkpoint engineering/[Applied Microbiology and Biotechnology]

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EID
272381
EOID
843058
Map
0
LastModified
2016年12月22日(木) 14:59:47
Operator
三木 ちひろ
Avail
TRUE
Censor
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Owner
鬼塚 正義
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種別 必須 学術論文(審査論文)
言語 必須 英語
招待 推奨
審査 推奨
カテゴリ 推奨
共著種別 推奨
学究種別 推奨
組織 推奨
  1. 徳島大学.工学部.生物工学科.生物機能工学講座
著者 必須
  1. (英) Kyoung Ho Lee
    役割 任意
    貢献度 任意
    学籍番号 推奨
  2. 鬼塚 正義([徳島大学.大学院社会産業理工学研究部.生物資源産業学域.応用生命系.生体分子機能学分野]/[徳島大学.生物資源産業学部.生物資源産業学科.応用生命講座])
    役割 任意
    貢献度 任意
    学籍番号 推奨
  3. (英) Honda Kohsuke
    役割 任意
    貢献度 任意
    学籍番号 推奨
  4. (英) Ohtake Hisao
    役割 任意
    貢献度 任意
    学籍番号 推奨
  5. 大政 健史
    役割 任意
    貢献度 任意
    学籍番号 推奨
題名 必須

(英) Rapid construction of transgene-amplified CHO cell lines by cell cycle checkpoint engineering

副題 任意
要約 任意

(英) The process of establishing high-producing cell lines for the manufacture of therapeutic proteins is usually both time-consuming and laborious due to the low probability of obtaining high-producing clones from a pool of transfected cells and slow cell growth under the strong selective pressure of screening to identify high-producing clones. We present a novel method to rapidly generate more high-producing cells by accelerating transgene amplification. A small interfering RNA (siRNA) expression vector against ataxia telangiectasia and Rad3 related (ATR), a cell cycle checkpoint kinase, was transfected into Chinese hamster ovary (CHO) cells. The influences of ATR downregulation on gene amplification and the productivity were investigated in CHO cells producing green fluorescent protein (GFP) and secreting monoclonal antibody (mAb). The ATR-downregulated cells showed up to a 6-fold higher ratio of GFP-positive cells than that of the control cell pool. Moreover, the downregulated mAb-producing cells had about a 4-fold higher specific production rate and a 3-fold higher volumetric productivity as compared with the mock cells. ATR-downregulated cells showed a much faster increase in transgene copy numbers during the gene amplification process via methotrexate (MTX) treatment in both GFP- and mAb-producing cells. Our results suggest that a pool of high-producing cells can be more rapidly generated by ATR downregulation as compared with conventional gene amplification by MTX treatment. This novel method may be a promising approach to reduce time and labor in the process of cell line development.

キーワード 推奨
  1. (英) Animals
  2. 生物工学(biotechnology)
  3. (英) CHO Cells
  4. (英) Cell Cycle Checkpoints
  5. (英) Cell Cycle Proteins
  6. (英) Cricetulus
  7. 遺伝子発現(gene expression)
  8. (英) Gene Silencing
  9. (英) Genes, Reporter
  10. 遺伝子工学(genetic engineering)
  11. (英) Green Fluorescent Proteins
  12. (英) Molecular Sequence Data
  13. (英) Recombinant Proteins
  14. (英) Sequence Analysis, DNA
  15. (英) Transgenes
発行所 推奨
誌名 必須 Applied Microbiology and Biotechnology([Springer-Verlag])
(pISSN: 0175-7598, eISSN: 1432-0614)
ISSN 任意 1432-0614
ISSN: 0175-7598 (pISSN: 0175-7598, eISSN: 1432-0614)
Title: Applied microbiology and biotechnology
Title(ISO): Appl Microbiol Biotechnol
Supplier: Springer Online Journal Archive
Publisher: Springer Berlin Heidelberg
 (NLM Catalog  (Scopus  (CrossRef (Scopus information is found. [need login])
必須 97
必須 13
必須 5731 5741
都市 任意
年月日 必須 2013年 4月 25日
URL 任意
DOI 任意 10.1007/s00253-013-4923-9    (→Scopusで検索)
PMID 任意 23615744    (→Scopusで検索)
NAID 任意
WOS 任意 000321121500008
Scopus 任意 2-s2.0-84879418366
評価値 任意
被引用数 任意
指導教員 推奨
備考 任意
  1. (英) Article.ELocationID: 10.1007/s00253-013-4923-9

  2. (英) Article.DataBankList.DataBank.DataBankName: GENBANK

  3. (英) Article.DataBankList.DataBank.AccessionNumberList.AccessionNumber: AB761036

  4. (英) Article.PublicationTypeList.PublicationType: Journal Article

  5. (英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't