| Autor: |
Permyakova, Natalya V., Sidorchuk, Yury V., Marenkova, Tatyana V., Khozeeva, Sofya A., Kuznetsov, Vitaly V., Zagorskaya, Alla A., Rozov, Sergei M., Deineko, Elena V. |
| Zdroj: |
Molecular Biology Reports; Dec2019, Vol. 46 Issue 6, p5735-5743, 9p |
| Abstrakt: |
Targeted genome editing using CRISPR/Cas9 is a promising technology successfully verified in various plant species; however, it has hardly been used in plant cell suspension cultures. Here, we describe a successful knockout of a green fluorescent protein (gfp) reporter gene in Arabidopsis cell culture. We transformed seven transgenic suspension cell lines carrying one to three gfp gene copies with a binary vector containing genes coding for Cas9 and guide RNAs targeting the gfp gene. We detected the site-specific mutations by restriction analysis of a gfp amplicon. DNA sequencing of the PCR products confirmed high diversity of insertion-deletion mutations in the cell lines after the editing. We also analyzed gfp mRNA expression by real-time PCR and observed a decrease in gfp transcription after the target site modification. We can conclude that the CRISPR/Cas9 system can be successfully used for introducing site-specific mutations into the genome of cultured suspension cells of Arabidopsis. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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