Development of rAAV-mediated method to modify common marmoset pre-implantation stage embryo genome

2013 Fiscal Year Final Research Report

• Project/Area Number: 23700521
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Laboratory animal science
• Research Institution: National Center of Neurology and Psychiatry
• Principal Investigator: OKADA Hironori 独立行政法人国立精神・神経医療研究センター, 神経研究所 遺伝子疾患治療研究部, 科研費研究員 (80416271)
• Project Period (FY): 2011 – 2013
• Keywords: アデノ随伴ウイルスベクター / コモンマーモセット / 受精卵 / 遺伝子挿入 / 疾患モデル動物

Research Abstract

Although generation of transgenic marmosets with random insertion of lentivirus vector genome into embryo was reported, the phenotypes of such an animal would be diversified depending on the integration site. To establish more advanced method, adeno-associated virus vector (rAAV)-mediated transduction into marmoset embryo, and inclusion of several gene-editing systems into rAAV were examined. The results were as follows: 1) Enhancement of rAAV2 genome integration into a marmoset cell line due to addition of rAAV5 expressing AAV2Rep. 2) Construction of AAV vector plasmid carrying TALE with phi-Gamma catalytic domain. 3) Induction of indel mutation to HEK293 cells by transfection of AAV vector plasmid carrying CRISPR/Cas9 system, and production of rAAV from the plasmid. 4) Efficient rAAV9-mediated transduction into marmoset embryo, and consequent robust transgene expression in ICM. These resulting objects and findings would be useful for production of genome-edited non-human primate.

Research Products

• [Journal Article] Effect of the size of zona pellucida opening on hatching in the common marmoset monkey (Callithrix jacchus) embryo (2013)
  • Author(s): Ishibashi H, Motohashi H, Kumon M, Yamamoto K, Okada H, Okada T, Seki K
  • Journal Title: Animal Science Journal Volume: Nov;84(11) Pages: 740-3
  • DOI: 10.1111/asj.12115
  • Peer Reviewed
• [Journal Article] Ultrasound-guided non-surgical embryo collection in the common marmoset (2013)
  • Author(s): Ishibashi H, Motohashi HH, Kumon M, Yamamoto K, Okada H, Okada T, Seki K
  • Journal Title: Reproductive Biology Volume: Jun;13(2) Pages: 139-44
  • DOI: 10.1016/j.repbio.2013.02.002
  • Peer Reviewed
• [Journal Article] Robust Long-term Transduction of Common Marmoset Neuromuscular Tissue With rAAV1 and rAAV9 (2013)
  • Author(s): Okada H, Ishibashi H, Hayashita-Kinoh H, Chiyo T, Nitahara-Kasahara Y, Baba Y, Watanabe S, Takeda S, Okada T
  • Journal Title: Molecular Therapy Nucleic Acids Volume: May 28;2 Pages: e95
  • DOI: 10.1038/mtna.2013.21
  • Peer Reviewed
• [Journal Article] Efficient Embryo Transfer in the Common Marmoset Monkey (Callithrix jacchus) with a Reduced Transfer Volume : A Non-Surgical Approach with Cryopreserved Late-Stage Embryos (2013)
  • Author(s): Ishibashi H, Motohashi HH, Kumon M, Yamamoto K, Okada H, Okada T, Seki K
  • Journal Title: Biology of Reproduction Volume: May 9;88(5) Pages: 115
  • DOI: 10.1095/biolreprod.113.109165
  • Peer Reviewed
• [Journal Article] Long-term Engraftment of Multipotent Mesenchymal Stromal Cells that Differentiate to Form Myogenic Cells in Dogs With Duchenne Muscular Dystrophy (2012)
  • Author(s): Nitahara-Kasahara Y, Hayashita-Kinoh H, Ohshima-Hosoyama S, Okada H, Wada-Maeda M, Nakamura A, Okada T, Takeda S
  • Journal Title: Molecular Therapy Volume: 20(1) Pages: 168-77
  • DOI: 10.1038/mt.2011.181
  • Peer Reviewed
• [Presentation] Generation of muscular dystrophy NHP model with rAAV 1 and 9-mediated transduction of common marmoset (2013)
  • Author(s): Okada H., Ishibashi H., Hayashita-Kinoh H., Chiyo T., Nitahara-Kasahara Y., Okada T., Takeda S.
  • Organizer: 19th Annual Meeting of JSGT, PS-6
  • Place of Presentation: Okayama 岡山コンベンションセンター
  • Year and Date: 20130704-06
• [Presentation] Effective Transduction of Common Marmoset with rAAV1 and 9 To Generate NHP Model of Muscular Dystrophy (2013)
  • Author(s): Hironori Okada, Hidetoshi Ishibashi, Hiromi Hayashita-Kinoh, Tomoko Chiyo, Yuko Nitahara-Kasahara, Takashi Okada, Shin'ichi Takeda
  • Organizer: 16th American Society of Gene & Cell Therapy
  • Place of Presentation: Salt lake city, USA Salt Palace Convention Center
  • Year and Date: 20130515-18
• [Presentation] Engraftment of Mesenchymal Stromal Cells That Can Differentiate To Form Myogenic Cells Is Enhanced by Expressing IL-10 in Dog with Duchenne Muscular Dystrophy (2013)
  • Author(s): Yuko N. Kasahara, Hiromi H. Kinoh, Tomoko Chiyo, Hironori Okada, Takashi Okada, Shin'ichi Takeda
  • Organizer: 16th American Society of Gene & Cell Therapy
  • Place of Presentation: Salt lake city, USA Salt Palace Convention Center
  • Year and Date: 20130515-18
• [Presentation] Immune Tolerance Induction in Canine X-Linked Muscular Dystrophy with Trans-Placental rAAV9-Microdystrophin Transduction (2013)
  • Author(s): Hiromi Hayashita-Kinoh, Hironori Okada, Yuko Nitahara-Kasahara, Tomoko Chiyo, Naoko Yugeta, Takashi Okada, Shin'ichi Takeda
  • Organizer: 16th American Society of Gene & Cell Therapy
  • Place of Presentation: Salt lake city, USA Salt Palace Convention Center
  • Year and Date: 20130515-18
• [Presentation] Effective transfer of morpholino into muscle cells by using AAV empty capsids (2012)
  • Author(s): Hayashita-Kinoh H., Okada H., Chiyo T., Nitahara-Kasahara Y., Okada T., Takeda S.
  • Organizer: 18th Annual Meeting of JSGT
  • Place of Presentation: Kumamoto ホテル熊本テルサ(PO-59)
  • Year and Date: 20120628-30
• [Presentation] Shakuyaku-kanzo-to promotes recombinant AAV cellular uptake to enhance transduction of cultured mammalian cells (2012)
  • Author(s): Chiyo T., Hayashita-Kinoh H., Nitahara-Kasahara Y., Okada H., Okada T., Takeda S.
  • Organizer: 18th Annual Meeting of JSGT
  • Place of Presentation: Kumamoto ホテル熊本テルサ(PO-57)
  • Year and Date: 20120628-30
• [Presentation] Effective transgene expression in non-human primate muscle with AAV type 9 vectors following immune suppression (2012)
  • Author(s): Ishii A., Okada H., Hayashita-Kinoh H., Shin J-H., Okada T., Takeda S.
  • Organizer: 18th Annual Meeting of JSGT
  • Place of Presentation: Kumamoto ホテル熊本テルサ(OR-31)
  • Year and Date: 20120628-30
• [Presentation] Strategy for efficient generation of DMD model marmoset with rAAV-mediated transduction (2012)
  • Author(s): Okada H., Ishibashi H., Hayashita-Kinoh H., Chiyo T., Nitahara-Kasahara Y., Okada T., Takeda S.
  • Organizer: 18th Annual Meeting of JSGT
  • Place of Presentation: Kumamoto ホテル熊本テルサ(OR-29)
  • Year and Date: 20120628-30
• [Presentation] rAAV8/9-Mediated Muscle Transduction with Tacrolimus in Non-Human Primate (2012)
  • Author(s): Akiko Ishii, Hironori Okada, Hiromi Hayashita-Kinou, Takashi Okada, Shin'ichi Takeda
  • Organizer: 15th American Society of Gene & Cell Therapy
  • Place of Presentation: Philadelphia, USA Pennsylvania Convention Center
  • Year and Date: 20120516-19
• [Presentation] rAAV9-Mediated Microdystrophin Gene Transfer with Immune Tolerance Induction Improves Dystrophic Phenotype of Canine X-Linked Muscular Dystrophy (2012)
  • Author(s): Hiromi Hayashita-Kinoh, Naoko Yugeta, Hironori Okada, Yuko Nitahara-Kasahara, Tomoko Chiyo, Takashi Okada, Shin'ichi Takeda
  • Organizer: 15th American Society of Gene & Cell Therapy
  • Place of Presentation: Philadelphia, USA Pennsylvania Convention Center
  • Year and Date: 20120516-19
• [Presentation] Strategy for rAAV-mediated transduction of common marmoset skeletal muscle to generate NHP DMD model (2012)
  • Author(s): Hironori Okada, Hidetoshi Ishibashi, Hiromi Hayashita-Kinoh, Tomoko Chiyo, Yuko Nitahara-Kasahara, Takashi Okada, and Shin'ichi Takeda
  • Organizer: 15th American Society of Gene & Cell Therapy
  • Place of Presentation: Philadelphia, USA Pennsylvania Convention Center
  • Year and Date: 20120516-19
• [Presentation] DISRUPTION OF COMMON MARMOSET DYSTROPHIN mRNA TO GENERATE NON-HUMAN PRIMATE DMD MODEL (2011)
  • Author(s): Hironori Okada, Hiromi Hayashita-Kinoh, Tomoko Chiyo, Hirohiko Hohjoh, Yuko Nitahara-Kasahara, Takashi Okada, Shin'ichi Takeda
  • Organizer: 17th Annual Meeting of JSGT
  • Place of Presentation: Fukuoka 九州大学医学部百年講堂(OR-46)
  • Year and Date: 20110715-17
• [Presentation] Effective transgene expression in non-human primate muscle with AAV type 9 vectors following immune suppression (2011)
  • Author(s): Ishii A., Okada H., Hayashita-Kinoh H., Shin J-H., Okada T., Takeda S.
  • Organizer: 17th Annual Meeting of JSGT
  • Place of Presentation: Fukuoka 九州大学医学部百年講堂(OR-63)
  • Year and Date: 20110715-17
• [Presentation] AAV empty capsids mediate effective nuclear transportation of morpholino in the muscle cells (2011)
  • Author(s): Hayashita-Kinoh H., Okada H., Chiyo T., Nitahara-Kasahara Y., Okada T., Takeda S.
  • Organizer: 17th Annual Meeting of JSGT
  • Place of Presentation: Fukuoka 九州大学医学部百年講堂(OR-64)
  • Year and Date: 20110715-17
• [Presentation] Immune Tolerance Induction in Canine X-Linked Muscular Dystrophy with rAAV9-Microdystrophin Transduction (2011)
  • Author(s): Hiromi Hayashita-Kinoh, Naoko Yugeta, Hironori Okada, Yuko Nitahara-Kasahara, Tomoko Chiyo, Takashi Okada, Shin'ichi Takeda
  • Organizer: 14th American Society of Gene & Cell Therapy
  • Place of Presentation: Seattle, USA Washington State Convention & Trade Center
  • Year and Date: 20110518-21
• [Presentation] Long-Term Engraftment of Mesenchymal Stromal Cells That Can Differentiate To Form Myogenic Cells in Dog with Duchenne Muscular Dystrophy (2011)
  • Author(s): Yuko Nitahara Kasahara, Hiromi Hayashita Kinoh, Hironori Okada, Jin-Hong Shin, Akiyo Nishiyama, Sachiko Ohshima Hosoyama, Michiko Wada Maeda, Akinori Nakamura, Takashi Okada, Shin'ichi Takeda
  • Organizer: 14th American Society of Gene & Cell Therapy
  • Place of Presentation: Seattle, USA Washington State Convention & Trade Center
  • Year and Date: 20110518-21