書誌事項
- タイトル別名
-
- Generation of DAT-integrin α5 heterozygous knock-in embryonic stem cells using CRISPR/Cas9 system
抄録
<p>We have previously found that integrin α5β1 on dopaminergic neurons plays an important role in the neurite outgrowth on striatal neurons. This finding indicates that integrin α5 (Itga)-overexpressing dopaminergic neurons enhance functional regeneration in transplantation therapy for Parkinson disease. Here, we generated the dopamine transporter (DAT)-Itga heterozygous knock-in mouse embryonic stem cells using the CRISPR)/Cas9 system. These cells can be induced to express Itga gene after dopaminergic differentiation, avoiding the loss of DAT function. The knock-in targeting vector expressing Venus (KI-Ctrl) or Itga followed by Venus (KI-Itga) contained a transgene of 2721 bp or 6470 bp, respectively, which was flanked by the 5'- and 3'- homology arms. Homology arms of approximately 2 kbp were required to obtain heterozygous recombinant clones. Although two KI-Ctrl cloned cells with accurate chromosomal sequence in non-targeted allele were obtained, all KI-Itga cloned cells had indel mutations. By decreasing the amount of Cas9-expressing plasmid and co-transfecting with the rescue vector containing chromosomal sequences, one KI-Itga5 cloned cells with accurate DNA sequence in non-targeted allele was obtained. </p>
収録刊行物
-
- 日本薬理学会年会要旨集
-
日本薬理学会年会要旨集 92 (0), 3-P-122-, 2019
公益社団法人 日本薬理学会
- Tweet
詳細情報 詳細情報について
-
- CRID
- 1390846609814535552
-
- NII論文ID
- 130007813318
-
- ISSN
- 24354953
-
- 本文言語コード
- ja
-
- データソース種別
-
- JaLC
- Crossref
- CiNii Articles
-
- 抄録ライセンスフラグ
- 使用不可