title: Tracking and transforming neocortical progenitors by CRISPR/Cas9 gene targeting and piggyBac transposase lineage labeling.
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.
authors.affiliation: Department of Physiology and Neurobiology, Institute for Systems Genomics, Institute for Brain and Cognitive Science, University of Connecticut, Storrs, CT 06268, USA.
authors.forename: Fuyi
authors.initials: F
authors.lastname: Chen
authors.affiliation: Department of Physiology and Neurobiology, Institute for Systems Genomics, Institute for Brain and Cognitive Science, University of Connecticut, Storrs, CT 06268, USA.
authors.forename: Joel
authors.initials: J
authors.lastname: Rosiene
authors.affiliation: Department of Physiology and Neurobiology, Institute for Systems Genomics, Institute for Brain and Cognitive Science, University of Connecticut, Storrs, CT 06268, USA.
authors.forename: Alicia
authors.initials: A
authors.lastname: Che
authors.affiliation: Department of Neuropathology, University of Bonn Medical Center, 53105 Born, Germany.
authors.forename: Albert
authors.initials: A
authors.lastname: Becker
authors.affiliation: Department of Physiology and Neurobiology, Institute for Systems Genomics, Institute for Brain and Cognitive Science, University of Connecticut, Storrs, CT 06268, USA Joseph.loturco@uconn.edu.
authors.forename: Joseph
authors.initials: J
authors.lastname: LoTurco
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology*; D001921:Brain / Q000378:metabolism; D001932:Brain Neoplasms / Q000473:pathology; D064113:CRISPR-Cas Systems / Q000235:genetics*; D019070:Cell Lineage; D022202:Chromosomes, Artificial, Bacterial; D018274:Electroporation; D005260:Female; D005909:Glioblastoma / Q000473:pathology; D006801:Humans; D016296:Mutagenesis; D009154:Mutation; D025542:Neurofibromin 1 / Q000378:metabolism; D009474:Neurons / Q000166:cytology; D051059:PTEN Phosphohydrolase / Q000378:metabolism; D011247:Pregnancy; D011270:Pregnancy, Animal; D051381:Rats; D017208:Rats, Wistar; D013234:Stem Cells / Q000166:cytology*; D019895:Transposases / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D025542:Neurofibromin 1; D016159:Tumor Suppressor Protein p53; D019895:Transposases; D051059:PTEN Phosphohydrolase
keywords: CRISPR/Cas9; Glioblastoma multiforme; In utero electroporation; Lineage; Neural progenitors; piggyBac