Theranostics 2020; 10(25):11507-11519. doi:10.7150/thno.46642 This issue Cite

Research Paper

Precise and efficient silencing of mutant KrasG12D by CRISPR-CasRx controls pancreatic cancer progression

Wang Jiang1,2,3,4*, Hao Li1,2,3,4*✉, Xiyu Liu5*, Jianping Zhang6, Wuhu zhang1,2,3,4, Tianjiao Li1,2,3,4, Liang Liu1,2,3,4✉, Xianjun Yu1,2,3,4✉

1. Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai 200032, PR China
2. Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, PR China
3. Shanghai Pancreatic Cancer Institute, Shanghai 200032, PR China
4. Pancreatic Cancer Institute, Fudan University, Shanghai 200032, PR China
5. Department of Breast Surgery, Fudan University Shanghai Cancer Center, Shanghai 200032, PR China
6. Department of Nuclear Medicine, Fudan University Shanghai Cancer Centre, Shanghai 200032, PR China
*These authors contributed equally to this study.

Citation:
Jiang W, Li H, Liu X, Zhang J, zhang W, Li T, Liu L, Yu X. Precise and efficient silencing of mutant KrasG12D by CRISPR-CasRx controls pancreatic cancer progression. Theranostics 2020; 10(25):11507-11519. doi:10.7150/thno.46642. https://www.thno.org/v10p11507.htm
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Abstract

Graphic abstract

Rationale: Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease with few therapeutic targets and rare effective treatments. Over 90% of PDAC tumors bear a Kras mutation, and the single-site mutation G12D (KrasG12D) is most prevalent.

Methods: Here, we applied the CRISPR-CasRx system to silence the mutant KrasG12D transcript in PDAC cells. We also used a capsid-optimized adenovirus-associated virus 8 vector (AAV8) to deliver the CRISPR-CasRx system into PDAC orthotopic tumors and patient-derived tumor xenografts (PDX).

Results: Our data showed that guided by a KrasG12D-specific gRNA, CasRx is able to precisely and efficiently silence the mutant KrasG12D expression in PDAC cells. The knockdown of mutant KrasG12D by CasRx abolishes the aberrant activation of downstream signaling induced by mutant KrasG12D and subsequently suppresses the tumor growth and improves the sensitivity of gemcitabine in PDAC. Additionally, delivering CasRx-gRNA via AAV8 into the orthotopic KrasG12D PDAC tumors substantially improves the survival of mice without obvious toxicity. Furthermore, targeting KrasG12D through CasRx suppresses the growth of PDAC PDXs. In conclusion, our study provides a proof-of-concept that CRISPR-CasRx can be utilized to target and silence mutant KrasG12D transcripts and therefore inhibit PDAC malignancy.

Keywords: PDAC, KrasG12D, CasRx, off-target, gRNA.


Citation styles

APA
Jiang, W., Li, H., Liu, X., Zhang, J., zhang, W., Li, T., Liu, L., Yu, X. (2020). Precise and efficient silencing of mutant KrasG12D by CRISPR-CasRx controls pancreatic cancer progression. Theranostics, 10(25), 11507-11519. https://doi.org/10.7150/thno.46642.

ACS
Jiang, W.; Li, H.; Liu, X.; Zhang, J.; zhang, W.; Li, T.; Liu, L.; Yu, X. Precise and efficient silencing of mutant KrasG12D by CRISPR-CasRx controls pancreatic cancer progression. Theranostics 2020, 10 (25), 11507-11519. DOI: 10.7150/thno.46642.

NLM
Jiang W, Li H, Liu X, Zhang J, zhang W, Li T, Liu L, Yu X. Precise and efficient silencing of mutant KrasG12D by CRISPR-CasRx controls pancreatic cancer progression. Theranostics 2020; 10(25):11507-11519. doi:10.7150/thno.46642. https://www.thno.org/v10p11507.htm

CSE
Jiang W, Li H, Liu X, Zhang J, zhang W, Li T, Liu L, Yu X. 2020. Precise and efficient silencing of mutant KrasG12D by CRISPR-CasRx controls pancreatic cancer progression. Theranostics. 10(25):11507-11519.

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