Inactivation of an integrated antibiotic resistance gene in mammalian cells to re-enable antibiotic selection
Removing an antibiotic resistance gene allows the same antibiotic to be re-used in the next round of genetic manipulation. Here we applied the CRISPR/Cas system to disrupt the puromycin resistance gene in an engineered mouse embryonic stem cell line and then re-used puromycin selection in the result...
| Publié dans: | BioTechniques. - 1991. - 56(2014), 4 vom: 14., Seite 198-201 |
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| Auteur principal: | |
| Autres auteurs: | , , |
| Format: | Article en ligne |
| Langue: | English |
| Publié: |
2014
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| Accès à la collection: | BioTechniques |
| Sujets: | Journal Article Research Support, Non-U.S. Gov't CRISPR/Cas antibiotic resistance gene genetic engineering Anti-Bacterial Agents Puromycin 4A6ZS6Q2CL |
| Résumé: | Removing an antibiotic resistance gene allows the same antibiotic to be re-used in the next round of genetic manipulation. Here we applied the CRISPR/Cas system to disrupt the puromycin resistance gene in an engineered mouse embryonic stem cell line and then re-used puromycin selection in the resulting cells to establish stable reporter cell lines. With the CRISPR/Cas system, pre-engineered sequences, such as loxP or FRT, are not required. Thus, this technique can be used to disrupt antibiotic resistance genes that cannot be removed by the Cre-loxP and Flp-FRT systems |
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| Description: | Date Completed 15.12.2014 Date Revised 14.04.2014 published: Electronic-eCollection Citation Status MEDLINE |
| ISSN: | 1940-9818 |
| DOI: | 10.2144/000114160 |