Multiplexed genome editing by in vivo electroporation of Cas9 ribonucleoproteins effectively induces endometrial carcinoma in mice

Synergistic effects among multiple gene mutations are involved in cancer development and progression. However, developing genetically modified mouse models to analyze various combinations of mutations is extremely labor‐intensive and time‐consuming. To address these problems, we developed a novel me...

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Bibliographic Details
Published in:International journal of cancer 2023-06, Vol.152 (11), p.2331-2337
Main Authors: Kobayashi, Ryosuke, Kawabata‐Iwakawa, Reika, Sugiyama, Makoto, Oyama, Tetsunari, Ohtsuka, Masato, Horii, Takuro, Morita, Sumiyo, Nishiyama, Masahiko, Hatada, Izuho
Format: Article
Language:English
Subjects:
Animal models
Animals
Cancer
Carcinogenesis
Carcinoma
CRISPR
CRISPR-Cas Systems
Electroporation
Electroporation - methods
Endometrial cancer
endometrial carcinoma
Endometrial Neoplasms - genetics
Endometrium
Epithelial cells
Female
Gene Editing - methods
Genome editing
Genomes
Humans
LKB1 protein
Medical research
Mice
mouse model
Mutation
PTEN protein
ribonucleoprotein complex
Ribonucleoproteins
Ribonucleoproteins - genetics
Uterine cancer
Uterus
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Summary:Synergistic effects among multiple gene mutations are involved in cancer development and progression. However, developing genetically modified mouse models to analyze various combinations of mutations is extremely labor‐intensive and time‐consuming. To address these problems, we developed a novel method for in vivo multiplexed genome editing of the murine uterus to model human endometrial carcinoma (EMC). To do this, we injected a CRISPR‐Cas9 ribonucleoprotein complex into the uterine cavity of adult female mice, followed by electroporation. Evaluation of reporter mice demonstrated that genome editing occurred specifically in uterine epithelial cells, which are the origin of EMCs. Simultaneous targeting of Pten/Trp53/Lkb1, or targeting of Pten/Lkb1 along with the Ctnnb1ΔEx3 mutation, resulted in efficient generation of invasive tumors in wild‐type females within 3 months. This novel method will enable rapid and easy validation of many combinations of gene mutations that lead to endometrial carcinogenesis. What's new? Synergy among multiple mutations influences the development of human endometrial carcinoma. Assessing combinations of mutations in conventional mouse models, however, is costly and time consuming. In the present study, to more feasibly reproduce multiple cancer mutations in mice, the authors experimented with in vivo genome editing via introduction of Cas9 ribonucleoproteins into the murine uterus. In wild‐type female mice, invasive tumors developed within 3 months of simultaneous introduction of mutations in Pten/Trp53/Lkb1 or Pten/Lkb1 and Ctnnb1 via genome editing. The novel method reported here enables rapid and simple validation of various combinations of mutations responsible for endometrial carcinogenesis.
ISSN:0020-7136
1097-0215
DOI:10.1002/ijc.34342