CRISPR-Cas9 Knockin Mice for Genome Editing and Cancer Modeling

Details

• Alternative Title:
  Cell
• Personal Author:
  Platt, Randall J. ; Chen, Sidi ; Zhou, Yang ; Yim, Michael J. ; Swiech, Lukasz ; Kempton, Hannah R. ; Dahlman, James E. ; Parnas, Oren ; Eisenhaure, Thomas M. ; Jovanovic, Marko ; Graham, Daniel B. ; Jhunjhunwala, Siddharth ; Xavier, Ramnik J. ; Langer, Robert ; Anderson, Daniel G. ; Hacohen, Nir ; Regev, Aviv ; Feng, Guoping ; Sharp, Phillip A. ; Zhang, Feng
• Description:
  CRISPR-Cas9 is a versatile genome editing technology for studying the functions of genetic elements. To broadly enable the application of Cas9 in vivo, we established a Cre-dependent Cas9 knockin mouse. We demonstrated in vivo as well as ex vivo genome editing using adeno-associated virus (AAV)-, lentivirus-, or particle-mediated delivery of guide RNA in neurons, immune cells, and endothelial cells. Using these mice, we simultaneously modeled the dynamics of KRAS, p53, and LKB1, the top three significantly mutated genes in lung adenocarcinoma. Delivery of a single AAV vector in the lung generated loss-of-function mutations in p53 and Lkb1, as well as homology-directed repair-mediated Kras(G12D) mutations, leading to macroscopic tumors of adenocarcinoma pathology. Together, these results suggest that Cas9 mice empower a wide range of biological and disease modeling applications.
• Subjects:
  Adenocarcinoma Animals Clustered Regularly Interspaced Short Palindromic Repeats Dendritic Cells Disease Models, Animal Gene Knock-In Techniques Genes, Tumor Suppressor Genetic Engineering Genetic Vectors Lentivirus Lung Neoplasms Mice Mice, Transgenic Oncogenes

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• Source:
  Cell. 2014; 159(2):440-455
• Pubmed ID:
  25263330
• Pubmed Central ID:
  PMC4265475
• Document Type:
  Journal Article
• Funding:
  P30 DK043351/DK/NIDDK NIH HHSUnited States/ ; HHMI/Howard Hughes Medical InstituteUnited States/ ; U54CA151884/CA/NCI NIH HHSUnited States/ ; P30-CA14051/CA/NCI NIH HHSUnited States/ ; P50 HG006193/HG/NHGRI NIH HHSUnited States/ ; R01 DK097768/DK/NIDDK NIH HHSUnited States/ ; R37 EB000244/EB/NIBIB NIH HHSUnited States/ ; EB000244/EB/NIBIB NIH HHSUnited States/ ; R01 EB000244/EB/NIBIB NIH HHSUnited States/ ; R01-CA133404/CA/NCI NIH HHSUnited States/ ; U54 CA151884/CA/NCI NIH HHSUnited States/ ; P30 CA014051/CA/NCI NIH HHSUnited States/ ; DP1-MH100706/DP/NCCDPHP CDC HHSUnited States/ ; DK43351/DK/NIDDK NIH HHSUnited States/ ; R01 CA133404/CA/NCI NIH HHSUnited States/ ; DP1 MH100706/MH/NIMH NIH HHSUnited States/ ; R01-NS 07312401/NS/NINDS NIH HHSUnited States/
• Volume:
  159
• Issue:
  2
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:82be2fb865db3c5cd21c6a0ea24d319d945a5ffcab10ed24c040cad1a17a72d4
• Download URL:
  https://stacks.cdc.gov/view/cdc/30201/cdc_30201_DS1.pdf
• File Type:
  [PDF - 3.28 MB ]