1. Tracking and transforming neocortical progenitors by CRISPR/Cas9 gene targeting and piggyBac transposase lineage labeling.
- Author
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Chen F, Rosiene J, Che A, Becker A, and LoTurco J
- Subjects
- Animals, Brain metabolism, Brain Neoplasms pathology, Cell Lineage, Electroporation, Female, Glioblastoma pathology, Humans, Mutagenesis, Mutation, Neurofibromin 1 metabolism, Neurons cytology, PTEN Phosphohydrolase metabolism, Pregnancy, Pregnancy, Animal, Rats, Rats, Wistar, Tumor Suppressor Protein p53 metabolism, Astrocytes cytology, CRISPR-Cas Systems genetics, Chromosomes, Artificial, Bacterial, Stem Cells cytology, Transposases genetics
- Abstract
The ability to induce targeted mutations in somatic cells in developing organisms and then track the fates of those cells is a powerful approach both for studying neural development and for modeling human disease. The CRISPR/Cas9 system allows for such targeted mutagenesis, and we therefore tested it in combination with a piggyBac transposase lineage labeling system to track the development of neocortical neural progenitors with targeted mutations in genes linked to neurodevelopmental disruptions and tumor formation. We show that sgRNAs designed to target PTEN successfully decreased PTEN expression, and led to neuronal hypertrophy and altered neuronal excitability. Targeting NF1, by contrast, caused increased astrocytogenesis at the expense of neurogenesis, and combined targeting of three tumor suppressors (PTEN, NF1 and P53) resulted in formation of glioblastoma tumors. Our results demonstrate that CRISPR/Cas9 combined with piggyBac transposase lineage labeling can produce unique models of neurodevelopmental disruption and tumors caused by somatic mutation in neural progenitors., (© 2015. Published by The Company of Biologists Ltd.)
- Published
- 2015
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