Generation of knock out rat using mouse <-> rat chimera

2014 Fiscal Year Final Research Report

• Project/Area Number: 24500490
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: Osaka University
• Principal Investigator: ISOTANI Ayako 大阪大学, 免疫学フロンティア研究センター, 特任准教授 (20444523)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 疾患モデル動物 / 遺伝子組換え / 異種キメラ

Outline of Final Research Achievements

In a previous study, we produced mouse←rat chimera by injecting rat embryonic stem (ES) cells into mouse blastocysts. In this model, rat ES cells contributed to various organs, including spermatozoa, indicating potential utility for the production of a knockout rat. Hence, we established new rat ES cell lines to visualize rat spermatozoa using green fluorescent protein (GFP) from green body-green sperm (GBGS) doubly transgenic rats. Subsequently, we prepared a homologous recombined rat ES cell lines. When GBGS-rat ES cells were used to produce mouse←rat chimera, rat spermatozoa were easily detected with GFP in chimeric testes. Subsequently, the germline potential of rat spermatozoa in the chimera were assessed by intracytoplasmic injections. 　 Rat pups were generated from rat spermatozoa in chimeric testes, and normal germline potential of rat spermatozoa in the chimeric testes was demonstrated. This study provides a novel method for the production of knockout rat.

Free Research Field

実験動物学