Targeted knock-in of an scFv-Fc antibody gene into the hprt locus of Chinese hamster ovary cells using CRISPR/Cas9 and CRIS-PITCh systems

Kawabe Yoshinori, Shinya Komatsu, Shodai Komatsu, Mai Murakami, Akira Ito, Tetsushi Sakuma, Takahiro Nakamura, Takashi Yamamoto, Masamichi Kamihira

研究成果: ジャーナルへの寄稿記事

8 引用 (Scopus)

抄録

Chinese hamster ovary (CHO) cells have been used as host cells for the production of pharmaceutical proteins. For the high and stable production of target proteins, the transgene should be integrated into a suitable genomic locus of host cells. Here, we generated knock-in CHO cells, in which transgene cassettes without a vector backbone sequence were integrated into the hprt locus of the CHO genome using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and CRISPR-mediated precise integration into target chromosome (CRIS-PITCh) systems. We investigated the efficiency of targeted knock-in of transgenes using these systems. As a practical example, we generated knock-in CHO cells producing an scFv-Fc antibody using the CRIS-PITCh system mediated by microhomology sequences for targeting. We found that the CRIS-PITCh system can facilitate targeted knock-in for CHO cell engineering.

元の言語英語
ページ(範囲)599-605
ページ数7
ジャーナルJournal of Bioscience and Bioengineering
125
発行部数5
DOI
出版物ステータス出版済み - 5 1 2018

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Clustered Regularly Interspaced Short Palindromic Repeats
Single-Chain Antibodies
Chromosomes
Cricetulus
Antibodies
Ovary
Genes
Cells
Transgenes
Cell engineering
Proteins
Drug products
Cell Engineering
Genome
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

これを引用

Targeted knock-in of an scFv-Fc antibody gene into the hprt locus of Chinese hamster ovary cells using CRISPR/Cas9 and CRIS-PITCh systems. / Yoshinori, Kawabe; Komatsu, Shinya; Komatsu, Shodai; Murakami, Mai; Ito, Akira; Sakuma, Tetsushi; Nakamura, Takahiro; Yamamoto, Takashi; Kamihira, Masamichi.

:: Journal of Bioscience and Bioengineering, 巻 125, 番号 5, 01.05.2018, p. 599-605.

研究成果: ジャーナルへの寄稿記事

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abstract = "Chinese hamster ovary (CHO) cells have been used as host cells for the production of pharmaceutical proteins. For the high and stable production of target proteins, the transgene should be integrated into a suitable genomic locus of host cells. Here, we generated knock-in CHO cells, in which transgene cassettes without a vector backbone sequence were integrated into the hprt locus of the CHO genome using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and CRISPR-mediated precise integration into target chromosome (CRIS-PITCh) systems. We investigated the efficiency of targeted knock-in of transgenes using these systems. As a practical example, we generated knock-in CHO cells producing an scFv-Fc antibody using the CRIS-PITCh system mediated by microhomology sequences for targeting. We found that the CRIS-PITCh system can facilitate targeted knock-in for CHO cell engineering.",
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AU - Yoshinori, Kawabe

AU - Komatsu, Shinya

AU - Komatsu, Shodai

AU - Murakami, Mai

AU - Ito, Akira

AU - Sakuma, Tetsushi

AU - Nakamura, Takahiro

AU - Yamamoto, Takashi

AU - Kamihira, Masamichi

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