網膜・角膜・脊髄・坐骨神経へのエレクトロポレーションによる遺伝子導入

NEPA21 Type III エレクトロポレーション装置

NEPA21エレクトロポレーション装置

Sピンセット電極

ｹｰﾌﾞﾙ･ﾌｯﾄｽｲｯﾁ

網膜・角膜・脊髄・坐骨神経へのエレクトロポレーションによる遺伝子導入

アプリケーション

マウス・ラット網膜へのIn Vivoエレクトロポレーション

 

  1. 網膜下へDNAの注入
  2. エレクトロポレーション

 

  1. ピンセット電極
  2. ラットをピンセット電極でホールド

様々な発生ステージで採取したラットの網膜へのエレクトロポレーション法による遺伝子導入。

図A：P0でpCAG-GFPを網膜に導入して、P21（a～c）で採取。 強膜側から撮影。

図B：P0でpCAG-GFPを網膜に導入して、以下のステージで採取。
P2（d～f）、P5（g～i）

Cepko laboratory, Department of Genetics and Howard Hughes Medical Institute, Harvard Medical School 松田孝彦先生 提供

文献

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      Ex vivo 網膜

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      Mol Ther Nucleic Acids . 2025 Jul 1;36(3):102616.

        In vivo 網膜

        dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

        Wang Q, Xu X, Chen S, Lu R, Li L, Lo CH, Liu Z, Ning K, Li T, Kowal TJ, Wang B, Hartnett ME, Wang S, Qi LS, Sun Y.

        Sci Adv. 2024 Dec 20;10(51):eadn7540.

          In vivo 網膜

          Optimization of HITI-Mediated Gene Insertion for Rhodopsin and Peripherin-2 in Mouse Rod Photoreceptors: Targeting Dominant Retinitis Pigmentosa

          Onishi A, Tsunekawa Y, Mandai M, Ishimaru A, Ohigashi Y, Sho J, Yasuda K, Suzuki K, Izpisua Belmonte JC, Matsuzaki F, Takahashi M.

          Invest Ophthalmol Vis Sci. 2024 Nov 18;65(13):38.

            In vivo 網膜

            SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP Manipulation

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            Cell Rep. 2019 Jun 25;27(13):4003-4012.e6.

              Ex vivo 脊髄

              Explant Culture of the Embryonic Mouse Spinal Cord and Gene Transfer by ex vivo Electroporation

              Kinoshita-Kawada M, Hasegawa H, Hongu T, Yanagi S, Kanaho Y, Masai I, Mishima T, Chen X, Tsuboi Y, Rao Y, Yuasa-Kawada J, Wu JY.

              Bio Protoc. 2019 Sep 20;9(18):e3373.

                Ex vivo 網膜

                Quantitative Analysis of the ThrbCRM1-centered Gene Regulatory Network

                Souferi B, Emerson MM.

                Biol Open. 2019 Apr 26;8(4):bio039115.

                  In vivo 脊髄

                  Convallatoxin Enhance the Ligand-Induced Mu-Opioid Receptor Endocytosis and Attenuate Morphine Antinociceptive Tolerance in Mice

                  Chao PK, Chang HF, Ou LC, Chuang JY, Lee PT, Chang WT, Chen SC, Ueng SH, Hsu JT, Tao PL, Law PY, Loh HH, Yeh SH.

                  Sci Rep. 2019 Feb 20;9(1):2405.

                    Ex Vivo 脊髄

                    A crucial role for Arf6 in the response of commissural axons to Slit

                    Kinoshita-Kawada M, Hasegawa H, Hongu T, Yanagi S, Kanaho Y, Masai I, Mishima T, Chen X, Tsuboi Y, Rao Y, Yuasa-Kawada J, Wu JY

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                      In vivo 坐骨神経

                      Molecular analysis of axonal-intrinsic and glial-associated co-regulation of axon degeneration

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                        In Vivo/Ex Vivo 網膜

                        Gene Transfer of Prolyl Hydroxylase Domain 2 Inhibits Hypoxia-inducible Angiogenesis in a Model of Choroidal Neovascularization.

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                          In Vivo 脊髄

                          Morphine drives internal ribosome entry site-mediated hnRNP K translation in neurons through opioid receptor-dependent signaling.

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                            坐骨神経

                            Schwann cell-derived exosomes enhance axonal regeneration in the peripheral nervous system.

                            Lopez-Verrilli MA, Picou F, Court FA.

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                              In Vivo/Ex Vivo 網膜

                              Requirement of retinoic acid receptor β for genipin derivative-induced optic nerve regeneration in adult rat retina.

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                                In Vivo 脊髄

                                CCL-1 in the spinal cord contributes to neuropathic pain induced by nerve injury.

                                Akimoto N, Honda K, Uta D, Beppu K, Ushijima Y, Matsuzaki Y, Nakashima S, Kido MA, Imoto K, Takano Y, Noda M.

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                                  Ex Vivo 網膜

                                  Identification of a Retina-Specific Otx2 Enhancer Element Active in Immature Developing Photoreceptors

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                                    Ex Vivo 網膜

                                    Bulk electroporation and population calcium imaging in the adult mammalian retina.

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                                      Ex Vivo 網膜

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                                        In Vivo/Ex Vivo 網膜

                                        A role for ligand-gated ion channels in rod photoreceptor development

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                                          In Vivo/Ex Vivo 網膜

                                          Electroporation and RNA interference in the rodent retina in vivo and in vitro

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                                            In Vivo/Ex Vivo 角膜

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                                              In Vivo/Ex Vivo 網膜

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                                              エレクトロポレーション

                                              ドラッグデリバリー・遺伝子導入

                                              電気式細胞融合

                                              蛍光組織染色・in situ HCR

                                              細胞凍結

                                              培養細胞 分離・灌流・計数・伸展・解析

                                              微生物 計数・画像解析・検査

                                              1細胞回収・マイクロダイセクション

                                              In Vivo イメージング・モニタリング

                                              卵振動培養