Manipulating gene expressions by electroporation in the developing brain of mammalian embryos.
One of the goals of developmental neuroscience in the post-genomic era is to clarify functions of a huge number of anonymous genes of which only DNA sequences are identified. More convenient methods for genetic manipulation in vertebrates, especially mammals, could help us to identify functions of the novel genes. Here we introduce a novel gene transfer technology using electroporation (EP), which is a simple and powerful strategy for genetic analysis. We have applied this method to cultured mammalian embryos in order to understand the function of specific genes in the developing brain. We have also performed EP in developing fetuses in utero guided by ultrasound image. The combination of these techniques in addition to analysis of genetic mutants will clarify functions of individual genes, gene interactions, and the molecular mechanisms underlying the brain development.
Takahashi
Masanori
M
Department of Developmental Neurobiology, Tohoku University Graduate School of Medicine, 2-1, Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
Sato
Kenichi
K
Nomura
Tadashi
T
Osumi
Noriko
N
D000818:Animals; D001921:Brain / Q000196:embryology* / Q000502:physiology*; D046508:Culture Techniques / Q000379:methods; D051959:ELAV Proteins; D018274:Electroporation / Q000379:methods*; D004622:Embryo, Mammalian; D005136:Eye Proteins; D005260:Female; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins / Q000187:drug effects / Q000235:genetics; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009154:Mutation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; D011247:Pregnancy; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D051381:Rats; D012097:Repressor Proteins; D012249:Rhombencephalon / Q000187:drug effects / Q000196:embryology / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics; D016216:Ultrasonography, Prenatal; D029961:Zebrafish Proteins
D051959:ELAV Proteins; D005136:Eye Proteins; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D016376:Oligonucleotides, Antisense; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; C000605733:Pax6 protein, rat; D016601:RNA-Binding Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins