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Targeting mutant KRAS with CRISPR-Cas9 controls tumor growth

Authors
 Wonjoo Kim  ;  Sangeun Lee  ;  Han Sang Kim  ;  Minjung Song  ;  Yong Hoon Cha  ;  Young-Hoon Kim  ;  Jeonghong Shin  ;  Eun-Seo Lee  ;  Yeonsoo Joo  ;  Jae J. Song  ;  Eun Ju Choi  ;  Jae W. Choi  ;  Jinu Lee  ;  Moonkyung Kang  ;  Jong In Yook  ;  Min Goo Lee  ;  Yeon-Soo Kim  ;  Soonmyung Paik  ;  Hyongbum (Henry) Kim 
Citation
 Genome Research, Vol.28(3) : 374-382, 2018 
Journal Title
 Genome Research 
ISSN
 1088-9051 
Issue Date
2018
Abstract
KRAS is the most frequently mutated oncogene in human tumors, and its activating mutations represent important therapeutic targets. The combination of Cas9 and guide RNA from the CRISPR-Cas system recognizes a specific DNA sequence and makes a double-strand break, which enables editing of the relevant genes. Here, we harnessed CRISPR to specifically target mutant KRAS alleles in cancer cells. We screened guide RNAs using a reporter system and validated them in cancer cells after lentiviral delivery of Cas9 and guide RNA. The survival, proliferation, and tumorigenicity of cancer cells in vitro and the growth of tumors in vivo were determined after delivery of Cas9 and guide RNA. We identified guide RNAs that efficiently target mutant KRAS without significant alterations of the wild-type allele. Doxycycline-inducible expression of this guide RNA in KRAS-mutant cancer cells transduced with a lentiviral vector encoding Cas9 disrupted the mutant KRAS gene, leading to inhibition of cancer cell proliferation both in vitro and in vivo. Intra-tumoral injection of lentivirus and adeno-associated virus expressing Cas9 and sgRNA suppressed tumor growth in vivo, albeit incompletely, in immunodeficient mice. Expression of Cas9 and the guide RNA in cells containing wild-type KRAS did not alter cell survival or proliferation either in vitro and in vivo. Our study provides a proof-of-concept that CRISPR can be utilized to target driver mutations of cancers in vitro and in vivo.
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DOI
10.1101/gr.223891.117
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
2. College of Dentistry (치과대학) > Dept. of Oral Pathology (구강병리학교실) > 1. Journal Papers
2. College of Dentistry (치과대학) > Dept. of Oral and Maxillofacial Surgery (구강악안면외과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Otorhinolaryngology (이비인후과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Han Sang(김한상) ORCID logo https://orcid.org/0000-0002-6504-9927
Kim, Hyongbum(김형범) ORCID logo https://orcid.org/0000-0002-4693-738X
Paik, Soon Myung(백순명) ORCID logo https://orcid.org/0000-0001-9688-6480
Song, Jae Jin(송재진) ORCID logo https://orcid.org/0000-0001-8183-9550
Yook, Jong In(육종인) ORCID logo https://orcid.org/0000-0002-7318-6112
Lee, Min Goo(이민구) ORCID logo https://orcid.org/0000-0001-7436-012X
Cha, Yong Hoon(차용훈) ORCID logo https://orcid.org/0000-0003-1761-3260
Choi, Eun Chang(최은창)
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/163253
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