@article{3872f58aeb3045f69462b7ec0ea6cca6,
title = "A CRISPR-Cas13a system for efficient and specific therapeutic targeting of mutant KRAS for pancreatic cancer treatment",
abstract = "Mutant KRAS is a known driver oncogene in pancreatic cancer. However, this protein remains an “undruggable” therapeutic target. Inhibiting mutated KRAS expression at the mRNA level is a potentially effective strategy. Recently, a novel CRISPR-Cas effector, Cas13a has been reported to specifically knock down mRNA expression under the guidance of a single CRISPR-RNA in mammalian cells. Here we demonstrate that the CRISPR-Cas13a system can be engineered for targeted therapy of mutant KRAS in pancreatic cancer. In initial screening, we show that the bacterial Cas13a protein and crRNA significantly knock down mutant KRAS mRNA expression, identifying a CRISPR-Cas13a system that can induce up to a 94% knockdown efficiency. Introducing a single mismatch into the crRNA-target duplex enabled the CRISPR-Cas13a system to specifically recognize KRAS-G12D mRNA with no detectable effects on wild-type KRAS mRNA. More importantly, CRISPR-Cas13a-mediated KRAS-G12D mRNA knockdown potently induced apoptosis in vitro and elicited marked tumor shrinkage in mice. Our work describes an optimization strategy for the development of a CRISPR-Cas13a system to affect efficient and specific knockdown of the oncogenic mRNA, establishing the CRISPR-Cas13a system as a flexible, targeted therapeutic tool.",
keywords = "CRISPR-Cas13a, KRAS, Oncogenic mutation, Pancreatic cancer, mRNA knockdown",
author = "Xiao Zhao and Liang Liu and Jiayan Lang and Keman Cheng and Yongwei Wang and Xueyan Li and Jian Shi and Yanli Wang and Guangjun Nie",
note = "Funding Information: No potential conflicts of interest were disclosed. This work was supported by the National Basic Research Plan of China ( 2018YFA020035 ), the Key Research Program of the Chinese Academy of Sciences ( KGZD-EW-T06 ), the Innovation Research Group of National Natural Science Foundation ( 11621505 ), the Chinese Postdoctoral Science Foundation ( 2017M610839 ), the Key Research Project of Frontier Science of the Chinese Academy of Sciences ( QYZDJ-SSW-SLH022 ), the National Postdoctoral Program for Innovative Talents ( BX201600042 ), the National Natural Science Foundation of China ( 51673051 , 91543127 , 31300822 , 31500814 , 91440201 and 31571335 ) and the Academy of Medical Sciences-Newton Advanced Fellowship, UK . Funding Information: No potential conflicts of interest were disclosed. This work was supported by the National Basic Research Plan of China (2018YFA020035), the Key Research Program of the Chinese Academy of Sciences (KGZD-EW-T06), the Innovation Research Group of National Natural Science Foundation (11621505), the Chinese Postdoctoral Science Foundation (2017M610839), the Key Research Project of Frontier Science of the Chinese Academy of Sciences (QYZDJ-SSW-SLH022), the National Postdoctoral Program for Innovative Talents (BX201600042), the National Natural Science Foundation of China (51673051, 91543127, 31300822, 31500814, 91440201 and 31571335) and the Academy of Medical Sciences-Newton Advanced Fellowship, UK. Publisher Copyright: {\textcopyright} 2018 Elsevier B.V. Copyright: Copyright 2018 Elsevier B.V., All rights reserved.",
year = "2018",
month = sep,
day = "1",
doi = "10.1016/j.canlet.2018.05.042",
language = "English (US)",
volume = "431",
pages = "171--181",
journal = "Cancer Letters",
issn = "0304-3835",
publisher = "Elsevier Ireland Ltd",
}