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このアイテムのファイル:
| ファイル | 記述 | サイズ | フォーマット | |
|---|---|---|---|---|
| j.xpro.2021.100965.pdf | 3.92 MB | Adobe PDF | 見る/開く |
| タイトル: | Optimized electroporation of CRISPR-Cas9/gRNA ribonucleoprotein complex for selection-free homologous recombination in human pluripotent stem cells |
| 著者: | Xu, Huaigeng Kita, Yuto Bang, Uikyu Gee, Peter Hotta, Akitsu    https://orcid.org/0000-0002-2619-7441 (unconfirmed) |
| 著者名の別形: | 北, 悠人 堀田, 秋津 |
| キーワード: | Cell isolation CRISPR Genetics Molecular Biology Stem Cells |
| 発行日: | 17-Dec-2021 |
| 出版者: | Elsevier BV |
| 誌名: | STAR Protocols |
| 巻: | 2 |
| 号: | 4 |
| 論文番号: | 100965 |
| 抄録: | Selection-free, scarless genome editing in human pluripotent stem cells (PSCs) by utilizing ribonucleoprotein (RNP) of CRISPR-Cas9 is a useful tool for a variety of applications. However, the process can be hampered by time-consuming subcloning steps and inefficient delivery of the RNP complex and ssDNA template. Here, we describe the optimized protocol to introduce a single nucleotide change or a loxP site insertion in feeder-free, xeno-free iPSCs by utilizing MaxCyte and 4D-Nucleofector electroporators. For complete details on the use and execution of this protocol, please refer to Kagita et al. (2021) and Xu et al. (2019). |
| 著作権等: | © 2021 The Author(s). This is an open access article under the Creative Commons Attribution 4.0 International license. |
| URI: | http://hdl.handle.net/2433/275835 |
| DOI(出版社版): | 10.1016/j.xpro.2021.100965 |
| PubMed ID: | 34825222 |
| 出現コレクション: | 学術雑誌掲載論文等 |
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