One-step generation of conditional and reversible gene knockouts
| Autor: | William C. Skarnes, Sajith Perera, Roxana C. Mustata, Katie Tremble, José C. R. Silva, Tiago Grego, Katie Andrews, Jihoon Kim, Juergen Fink, Alessandra Merenda, Bon-Kyoung Koo, Amanda Andersson-Rolf |
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| Přispěvatelé: | Rebelo Da Silva, Jose [0000-0001-5487-1117], Apollo - University of Cambridge Repository |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Biology Biochemistry Article Cell Line Gene Knockout Techniques Mice 03 medical and health sciences Animals Humans Clustered Regularly Interspaced Short Palindromic Repeats Allele Molecular Biology Gene health care economics and organizations Embryonic Stem Cells beta Catenin Gene knockout Gene Editing Genetics Genome Cell Biology 3. Good health enzymes and coenzymes (carbohydrates) HEK293 Cells 030104 developmental biology embryonic structures CRISPR-Cas Systems Biotechnology |
| Zdroj: | Nature methods |
| ISSN: | 1548-7105 1548-7091 |
| DOI: | 10.1038/nmeth.4156 |
| Popis: | Loss-of-function studies are key for investigating gene function, and CRISPR technology has made genome editing widely accessible in model organisms and cells. However, conditional gene inactivation in diploid cells is still difficult to achieve. Here, we present CRISPR-FLIP, a strategy that provides an efficient, rapid and scalable method for biallelic conditional gene knockouts in diploid or aneuploid cells, such as pluripotent stem cells, 3D organoids and cell lines, by co-delivery of CRISPR-Cas9 and a universal conditional intronic cassette. |
| Databáze: | OpenAIRE |
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