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In vivo multiplex gene targeting with Streptococcus pyogens and Campylobacter jejuni Cas9 for pancreatic cancer modeling in wild-type animal

Authors
 Yoo Jin Chang  ;  Jihyeon Bae  ;  Yang Zhao  ;  Geonseong Lee  ;  Jeongpil Han  ;  Yoon Hoo Lee  ;  Ok Jae Koo  ;  Sunmin Seo  ;  Yang Kyu Choi  ;  Su Cheong Yeom 
Citation
 JOURNAL OF VETERINARY SCIENCE, Vol.21(2) : e26, 2020-03 
Journal Title
JOURNAL OF VETERINARY SCIENCE
ISSN
 1229-845X 
Issue Date
2020-03
MeSH
Animals ; CRISPR-Cas Systems* ; Campylobacter jejuni / genetics* ; Dependovirus / genetics ; Disease Models, Animal ; Gene Targeting* ; Mice ; Mice, Transgenic ; Pancreatic Neoplasms / microbiology* ; Streptococcus pyogenes / genetics*
Keywords
associated virus ; CRISPR ; gene editing ; Pancreatic cancer
Abstract
Pancreatic ductal adenocarcinoma is a lethal cancer type that is associated with multiple gene mutations in somatic cells. Genetically engineered mouse is hardly applicable for developing a pancreatic cancer model, and the xenograft model poses a limitation in the reflection of early stage pancreatic cancer. Thus, in vivo somatic cell gene engineering with clustered regularly interspaced short palindromic repeats is drawing increasing attention for generating an animal model of pancreatic cancer. In this study, we selected Kras, Trp53, Ink4a, Smad4, and Brca2 as target genes, and applied Campylobacter jejuni Cas9 (CjCas9) and Streptococcus pyogens Cas9 (SpCas9) for developing pancreatic cancer using adeno associated virus (AAV) transduction. After confirming multifocal and diffuse transduction of AAV2, we generated SpCas9 overexpression mice, which exhibited high double-strand DNA breakage (DSB) in target genes and pancreatic intraepithelial neoplasia (PanIN) lesions with two AAV transductions; however, wild-type (WT) mice with three AAV transductions did not develop PanIN. Furthermore, small-sized Cjcas9 was applied to WT mice with two AAV system, which, in addition, developed high extensive DSB and PanIN lesions. Histological changes and expression of cancer markers such as Ki67, cytokeratin, Mucin5a, alpha smooth muscle actin in duct and islet cells were observed. In addition, the study revealed several findings such as 1) multiple DSB potential of AAV-CjCas9, 2) peri-ductal lymphocyte infiltration, 3) multi-focal cancer marker expression, and 4) requirement of > 12 months for initiation of PanIN in AAV mediated targeting. In this study, we present a useful tool for in vivo cancer modeling that would be applicable for other disease models as well.
Files in This Item:
T9992020417.pdf Download
DOI
10.4142/jvs.2020.21.e26
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/190192
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