title: Supernova: A Versatile Vector System for Single-Cell Labeling and Gene Function Studies in vivo.
abstract: Here we describe "Supernova" series of vector systems that enable single-cell labeling and labeled cell-specific gene manipulation, when introduced by in utero electroporation (IUE) or adeno-associated virus (AAV)-mediated gene delivery. In Supernova, sparse labeling relies on low TRE leakage. In a small population of cells with over-threshold leakage, initial tTA-independent weak expression is enhanced by tTA/TRE-positive feedback along with a site-specific recombination system (e.g., Cre/loxP, Flpe/FRT). Sparse and bright labeling by Supernova with little background enables the visualization of the morphological details of individual neurons in densely packed brain areas such as the cortex and hippocampus, both during development and in adulthood. Sparseness levels are adjustable. Labeled cell-specific gene knockout was accomplished by introducing Cre/loxP-based Supernova vectors into floxed mice. Furthermore, by combining with RNAi, TALEN, and CRISPR/Cas9 technologies, IUE-based Supernova achieved labeled cell-specific gene knockdown and editing/knockout without requiring genetically altered mice. Thus, Supernova system is highly extensible and widely applicable for single-cell analyses in complex organs, such as the mammalian brain.
authors: Division of Neurogenetics, National Institute of Genetics, Mishima, 411-8540, Japan.|Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies), Mishima, 411-8540, Japan.
authors: Wenshu
authors: W
authors: Luo
authors: Division of Neurogenetics, National Institute of Genetics, Mishima, 411-8540, Japan.|Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies), Mishima, 411-8540, Japan.
authors: Hidenobu
authors: H
authors: Mizuno
authors: Division of Neurogenetics, National Institute of Genetics, Mishima, 411-8540, Japan.|Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies), Mishima, 411-8540, Japan.
authors: Ryohei
authors: R
authors: Iwata
authors: Division of Neurogenetics, National Institute of Genetics, Mishima, 411-8540, Japan.|Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies), Mishima, 411-8540, Japan.
authors: Shingo
authors: S
authors: Nakazawa
authors: Laboratory for Behavioral Genetics, RIKEN Brain Science Institute, Wako, 351-0198, Japan.
authors: Kosuke
authors: K
authors: Yasuda
authors: Laboratory for Behavioral Genetics, RIKEN Brain Science Institute, Wako, 351-0198, Japan.
authors: Shigeyoshi
authors: S
authors: Itohara
authors: Division of Neurogenetics, National Institute of Genetics, Mishima, 411-8540, Japan.|Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies), Mishima, 411-8540, Japan.
authors: Takuji
authors: T
authors: Iwasato
mesh_terms: D000818:Animals; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics; D018274:Electroporation / Q000379:methods; D005260:Female; D055785:Gene Knockdown Techniques / Q000379:methods; D055786:Gene Knockout Techniques / Q000379:methods; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D006624:Hippocampus / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D009474:Neurons / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics; D059010:Single-Cell Analysis / Q000379:methods