Abstract
CRISPR-Cas is an adaptive and heritable prokaryotic immune system which targets invasive genetic elements such as viruses and plasmids. In recent years it has been adapted for the site-specific manipulation of eukaryotic genomes by deleting or exchanging gene copies. The nuclease Cas9 recognizes a given sequence within a host cell by a complementary guide-RNA and inserts a double strand break which is repaired by the cell's own non-homologous recombination repair. Due to this unprecise repair mechanism mistakes are inserted and the gene is disrupted. In our experiment we destruct the gene of a viral thymidine kinase stably inserted into the genome of a human glioblastoma cell line. As the thymidine kinase gene is fused to the gene of Green Fluorescent Protein, the success of the experiment can be monitored by a disappearance of the green fluorescence. At the same time cells producing Cas9 show an orange-red fluorescence, because Cas9 is always produced together with the Orange Fluorescent Protein. Green will become red
| Item Type: | Journal article |
|---|---|
| Faculties: | Biology > Department Biology I Medicine |
| Subjects: | 500 Science > 570 Life sciences; biology 600 Technology > 610 Medicine and health |
| ISSN: | 0009-2851 |
| Language: | German |
| Item ID: | 54519 |
| Date Deposited: | 14. Jun 2018, 09:56 |
| Last Modified: | 04. Nov 2020, 13:34 |
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