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Generation of knockout mouse models of cyclin-dependent kinase inhibitors by engineered nuclease-mediated genome editing

Laboratory Animal Research volume 34, pages 264–269 (2018)Cite this article

Abstract

Cell cycle dysfunction can cause severe diseases, including neurodegenerative disease and cancer. Mutations in cyclin-dependent kinase inhibitors controlling the G1 phase of the cell cycle are prevalent in various cancers. Mice lacking the tumor suppressors p16Ink4a (Cdkn2a, cyclin-dependent kinase inhibitor 2a), p19Arf (an alternative reading frame product of Cdkn2a,), and p27Kip1 (Cdkn1b, cyclin-dependent kinase inhibitor 1b) result in malignant progression of epithelial cancers, sarcomas, and melanomas, respectively. Here, we generated knockout mouse models for each of these three cyclin-dependent kinase inhibitors using engineered nucleases. The p16Ink4a and p19Arf knockout mice were generated via transcription activator-like effector nucleases (TALENs), and p27Kip1 knockout mice via clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRISPR/Cas9). These gene editing technologies were targeted to the first exon of each gene, to induce frameshifts producing premature termination codons. Unlike preexisting embryonic stem cell-based knockout mice, our mouse models are free from selectable markers or other external gene insertions, permitting more precise study of cell cycle-related diseases without confounding influences of foreign DNA.

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Authors and Affiliations

  1. Department of Biochemistry, College of Life Science & Biotechnology and Yonsei Laboratory Animal Research Center, Yonsei University, Seoul, 03722, Korea

    Bo Min Park, Jae-il Roh, Jaehoon Lee & Han-Woong Lee

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  1. Bo Min Park
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  2. Jae-il Roh
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  4. Han-Woong Lee
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Correspondence to Jae-il Roh.

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Park, B.M., Roh, Ji., Lee, J. et al. Generation of knockout mouse models of cyclin-dependent kinase inhibitors by engineered nuclease-mediated genome editing. Lab Anim Res 34, 264–269 (2018). https://doi.org/10.5625/lar.2018.34.4.264

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  • DOI: https://doi.org/10.5625/lar.2018.34.4.264

Keywords

  • TALEN
  • CRISPR/Cas9
  • cyclin-dependent kinase inhibitor

Laboratory Animal Research

ISSN: 2233-7660