Cre-loxP系マウス着床前発生におけるレポーター遺伝子の細胞系譜特異的発現を可能にする

スポンサーリンク

概要

• 論文の詳細を見る

• The Cre-loxP site-specific recombination system is useful for tracing the fate of cells via transient expression of the Cre recombinase gene directly introduced into the cells with a vector carrying loxP sites. In this study, we used this system to examine whether cell lineage-specific expression of a reporter gene occurs in murine preimplantation embryos. An expression plasmid, pCETZ-17, which consists of cytomegalovirus enhancer/chicken β-actin promoter (CAG), a portion of the rabbit β-globin gene (including the 2nd intron, 3rd exon and 3 noncoding region), loxP-flanked DNA sequence [containing enhanced green fluorescent protein (EGFP) cDNA and chloramphenicol acetyltransferase gene (CAT)], and lacZ gene encoding E. coli β-galactosidase (β-gal), was constructed. When pCETZ-17 DNA was microinjected into the pronuclei of fertilized eggs, 30.0% (6/20) of the surviving 2-cell embryos exhibited distinct fluorescence in both blastomeres, but never expressed lacZ protein, as evaluated by histochemical staining with X-Gal, a substrate for β-gal. When both plasmids, pCETZ-17 and pCAG/NCre (containing CAG and DNA sequences encoding nuclear location signal and Cre), were co-injected into fertilized eggs, all (12/12) embryos exhibited low or no fluorescence, but 66.7% (8/12) exhibited positive staining for β-gal activity in one or both blastomeres. This indicates that transient expression of Cre removed the loxP-flanked DNA sequence in pCETZ-17 and then caused expression of the downstream lacZ sequence. We next microinjected pCETZ-17 into the pronuclei of fertilized eggs, cultured these injected eggs for 1 day, and collected only 2-cell embryos expressing EGFP in both blastomeres. One blastomere of the EGFP-expressing 2-cell embryos was microinjected with pCAG/NCre, and these treated embryos were cultured for 2 days up to 8-cell stage. When the developing 8-cell embryos were subjected to staining with X-Gal, cell lineage-related staining pattern for β-gal activity was observed in all (7/7) embryos. These findings indicate that this Cre-loxP system, which is based on transient expression of the Cre gene directly introduced into nuclei of embryonic cells by microinjection, is useful for tracing cell lineage as well as for expressing a gene of interest in a lineage-specific manner in early embryogenesis

• 日本繁殖生物学会の論文

著者

• SATO Masahiro

  Department of Nuclear Engineering, School of Engineering, Tokai University

• MIYAZAKI Jun-ichi

  Department of Disease-related Gene Regulation Research (Sandoz)

• WATANABE Toshiteru

  Department of Pediatirics, School of Medicine, Tokai University

• KIMURA MINORU

  Department of DNA Biology, Tokai University School of Medicine

• YASUOKA Yukiko

  Department of Physiology, Kitasato University School of Medicine

• Kodama Hisako

  Department of Molecular and Developmental Science, Molecular Medicine Research Center, The Institute

関連論文

• Optimization of a real-time PCR assay to quantitate airborne fungi collected on a gelatin filter(METHODS)
• レｰザｰ誘起蛍光法(LIFS)によるプラズマ中の酸素ラジカル測定
• 磁化シｰトプラズマを用いたガス･ダイバｰタの基礎実験
• Accumulation of cholesterol and G_ ganglioside in cells cultured in the presence of progesterone:an implication for the basic defect in Niemann-Pick disease typc C
• Clonal prevalence of T cells infiltrating into the pancreas of prediabetic non-obese diabetic mice
• A Trial of Alloreactive T-cell Depletion Using Biotinylated Galactose Oxidase for the Prevention of Acute Graft-versus-Host Diseases
• Defective IL-5-receptor-mediated signaling in B cells of X-linked immunodeficient mice
• Establishment of a Mouse Macula Densa Cell Line with an nNOS Promoter Driving EGFP Expression
• IN SITU LOCALIZATION OF c-MYC mRNA IN HL-60 CELLS USING NON-RADIOACTIVE SYNTHETIC OLIGODEOXYNUCLEOTIDE PROBES
• Nitric Oxide Is lmportant for Mouse Beta-Cell Line Killing by Peritoneal Exudate Cells Obtained from Cyclophosphamide Treated Non-Obese Diabeitc Mice
• Structure of Demethylallosamidin as an Insect Chitinase Inhibitor(Organic Chemistry)
• Rapid Screening of a Novel Arrayed Medaka (Oryzias latipes) Cosmid Library
• Folliculogenesis following syngeneic transplantation of young murine ovaries into the testes
• Application of Extended Finite-Differernce Time-Domain Method to Two-Dimensional Dynamic Analysis of a Piezoelectric Vibrator
• Localization of the Sulphonylurea Receptor Subunits, SUR2A and SUR2B, in Rat Renal Tubular Epithelium
• PJ-119 Effect of Anti-platelet Drugs at Angiogenic Cell Therapy(Peripheral circulation-8 (H) PJ20,Poster Session (Japanese),The 70th Anniversary Annual Scientific Meeting of the Japanese Circulation Society)
• Suppression-burst patterns in intractable epilepsy with focal cortical dysplasia
• THE PHYLOGEOGRAPHY FOR CONSERVATION OF THE GENUS LEFUA(Taxonomy and Systematics,Abstracts of papers presented at the 75^ Annual Meeting of the Zoological Society of Japan)
• 光脱離法によるシートプラズマ中での負イオン密度の測定
• DNA immunization with Plasmodium falciparum serine repeat antigen: regulation of humoral immune response by coinculation of cytokine expression plasmid
• Suppression of T-h1 cell activation and prevention of autoimmune diabetes in NOD mice by local expression of viral IL-10
• Nervous diseases and Kampo(Japanese herbal)medicine:a new paradigm of therapy against intractable nervous diseases
• Preferential requirement of CD3ζ-mediated signals for development of immature rather than mature thymocytes
• Cre-loxP系マウス着床前発生におけるレポーター遺伝子の細胞系譜特異的発現を可能にする
• Fine Structures of Proton Resonance Spectra of Fluorobenzene Derivatives
• Critical Phenomena of the S=1/2 Heisenberg-Ising Model Studied by CVM and CAM
• Thermodynamics and Electrodynamics of Superradiant Phase
• Two-Time Green's Function Theory for Laser
• Comparison of dry- and wet-based fine bead homogenizations to extract DNA from fungal spores(METHODS)
• Monitoring and switching of cortico-basal ganglia loop functions by the thalamo-striatal system
• Instabilities and Possible Phases of Quasi-One-Dimensional Polyacenic Semiconductors : Progress Letters
• Anomalies in Transgenic Mice Carrying the Human Interleukin-2 Gene
• A Surgical Case for a Carbomedics Stuck Valve Due to Thrombotic Pannus Formation in the Mitral Position
• Nonconvulsive Status Epilepticus in Childhood Localization-Related Epilepsy
• Identification of the epileptic focus by the dipole tracing method : Single layered model vs Scalp-Skull-Brain model
• Renormalization-Group Theory for Quasi-One-Dimensional Conductor : Condensed Matter and Statistical Physics
• Possible Phases and Some Properties of the One-Dimensional Metal with the Half-Filled Band
• On the Possible Phases of a One-Dimensional Metal with a Half-Filled Band
• EVALUATION OF THE TYPE V PIT PATTERN IN THE LESIONS OF COLONIC Tis AND T1 CANCER
• Experience with an Enzyme-Linked Immuno Sorbent Assay for the Quantitation of Urinary Desmosine
• Management of transvaginal ultrasound-guided absolute ethanol sclerotherapy for ovarian endometriotic cysts
• Benign endometrial adenofibroma and polyp in patients receiving tamoxifen : findings on transvaginal ultrasonography and magnetic resonance imaging
• Cre-loxP System Confers Cell Lineage-Specific Expression of a Reporter Gene in Murine Preimplantation Development

もっと見る 閉じる

スポンサーリンク