Generation of knockout mouse models of cyclin-dependent kinase inhibitors by engineered nuclease-mediated genome editing.
10.5625/lar.2018.34.4.264
- Author:
Bo Min PARK
1
;
Jae il ROH
;
Jaehoon LEE
;
Han Woong LEE
Author Information
1. Department of Biochemistry, College of Life Science & Biotechnology, Yonsei University, Seoul, Korea. hwl@yonsei.ac.kr jhlee13@gmail.com rohjaeil@gmail.com
- Publication Type:Original Article
- Keywords:
TALEN;
CRISPR/Cas9;
cyclin-dependent kinase inhibitor
- MeSH:
Animals;
Cell Cycle;
Codon, Nonsense;
Cyclin-Dependent Kinase Inhibitor p16;
DNA;
Exons;
G1 Phase;
Genome*;
Melanoma;
Mice;
Mice, Knockout*;
Mutagenesis, Insertional;
Neurodegenerative Diseases;
Phosphotransferases*;
Reading Frames;
Sarcoma
- From:Laboratory Animal Research
2018;34(4):264-269
- CountryRepublic of Korea
- Language:English
-
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 p16(Ink4a) (Cdkn2a, cyclin-dependent kinase inhibitor 2a), p19(Arf) (an alternative reading frame product of Cdkn2a,), and p27(Kip1) (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 p16(Ink4a) and p19(Arf) knockout mice were generated via transcription activator-like effector nucleases (TALENs), and p27(Kip1) 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.
