CRISPR-Cas9 Knockin Mice for Genome Editing and Cancer Modeling
Open accessAbstract
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.
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Delivery of next-generation CRISPR tools by lentivirus or ribonucleoprotein complexes may be a suitable alternative strategy to DNA mutations in adult stem cells.
“Thus, delivery of next-generation CRISPR tools by lentivirus or ribonucleoprotein complexes may be a suitable alternative strategy to DNA mutations in adult stem cells.54,55,56”
From Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools (2024) · cites this paper as [56]
The Results section on ex vivo genome editing reports that lentiviral delivery of guide RNAs produced mutations in primary mouse dendritic cells. It does not establish ribonucleoprotein delivery or the proposed strategy in adult stem cells, nor does it describe genome editing as an alternative to DNA mutations. Page numbers are unavailable in our copy. (LLM, conf=0.86)
Experimental — accuracy not yet measuredCas9-dependent acute deletion of the Spi1 locus can be fast and highly efficient due to availability of Cas9-transgenic mice and vectors for high-level, synchronous guide RNA expression.
“Cas9-dependent acute deletion of the Spi1 locus can be fast and highly efficient due to the availability of Cas9-transgenic mice (92) and vectors that can be used for high-level, synchronous expression of guide RNAs.”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [92]
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- Authors
- Platt, R. J., Chen, S., Zhou, Y., Yim, M. J., Swiech, L., Kempton, H. R., Dahlman, J. E., Parnas, O., Eisenhaure, T. M., Jovanovic, M., Graham, D. B., Jhunjhunwala, S., Heidenreich, M., Xavier, R. J., Langer, R., Anderson, D. G., Hacohen, N., Regev, A., Feng, G., Sharp, P. A., & Zhang, F.
- Affiliations
- Broad Institute (US)
- Koch Institute for Integrative Cancer Research At MIT (US)
- McGovern Institute for Brain Research (US)
- Harvard–MIT Division of Health Sciences and Technology (US)
- Corresponding author
- Sharp, Phillip A.
- Zhang, Feng
- ORCID
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2014-10
- Volume
- 159
- Issue
- 2
- Pages
- 440-455
- ISSN
- 0092-8674 (ISSN-L)
- PMC
- PMC4265475
- OpenAlex
- W1972476556
- Cited by
- 2,001
- References
- 70
- Citations by year
- 2026: 121 · 2025: 162 · 2024: 191 · 2023: 188 · 2022: 202 · 2021: 179 · 2020: 190 · 2019: 171 · 2018: 175 · 2017: 159 · 2016: 140 · 2015: 108 · 2014: 9 · 2013: 1
- FWCI
- 56.82
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc-nd)
- PMC
- Funders
- Howard Hughes Medical Institute
- Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology
- National Institutes of Health (4r37eb000244-34)
- National Institute of Mental Health (DP1MH100706)
- National Human Genome Research Institute (P50HG006193)
- National Cancer Institute (R01CA133404, U54CA151884, P30CA014051, P30-CA-14051)
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK097768, DK 43351, P30DK043351)
- National Institute of Neurological Disorders and Stroke (R01NS073124)
- National Institute of Biomedical Imaging and Bioengineering (EB-000244, R37-EB000244)
- National Center for Chronic Disease Prevention and Health Promotion (DP1‑MH100706)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- CRISPR/Cas9, knock-in mice, in vivo gene editing, adeno-associated virus vectors, guide RNA delivery, lung adenocarcinoma, KRAS G12D, p53, LKB1, tumor modeling, homology-directed repair, neurons, immune cells, endothelial cells
- MeSH terms
- Adenocarcinoma, Animals, Dendritic Cells, Disease Models, Animal, Genetic Engineering, Genetic Vectors, Lung Neoplasms, Mice, Transgenic, Oncogenes, Lentivirus, Genes, Tumor Suppressor, Mice, Gene Knock-In Techniques, Clustered Regularly Interspaced Short Palindromic Repeats
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