| Autor: |
Hanako, Sato-Yazawa, Korehito, Kashiwagi, Jun, Ishii, Meitetsu, Masawa, Masato, Onozaki, Chie, Miyata-Hiramatsu, Tadasuke, Miyazawa, Mai, Sakaguchi, Takuya, Yazawa |
| Jazyk: |
angličtina |
| Rok vydání: |
2021 |
| Předmět: |
|
| Zdroj: |
Dokkyo Journal of Medical Sciences. 48(2):75-80 |
| ISSN: |
0385-5023 |
| Popis: |
In recent decades, genetic engineering and its application to medical sciences has been rapidly progressing, and gene transfection and genome-editing techniques are generally used even in medical investigations. Although establishing single-cell clones are essential for confirming the cellular phenotypic changes in association with genetic modifications, it is technically hard to dispense single-cell efficiently and correctly. Here, we demonstrate single-cell cloning of genetically modified lung cancer cells using a single particle isolation system, On-chip SPiS, by which bi-allelic gene-knockout cells and highly gene-expressing transfectants can be easily and certainly established. In our experiment, we obtained one bi-allelic knockout clone from only 12 clones（8.3％）and eight highly transgene-expressing clones from 20 clones（40％）. Furthermore, we could define that considerable rate of clones without transgene were mixed among selection marker-resistant cell populations. These results suggest that the single-cell dispensing system is a quite useful tool for single-cell cloning in genome-editing and gene-transfection experiments. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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