Genome Editing of the SNAI1 Gene in Rhabdomyosarcoma: A Novel Model for Studies of Its Role
| Autor: | Claudio Mussolino, Aleksandra Ulman, Toni Cathomen, Marcin Majka, Klaudia Skrzypek, Paweł Konieczny |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Gene Expression Histone Deacetylase 1 SNAI1 Gene Computational biology Biology Article 03 medical and health sciences 0302 clinical medicine Genome editing TALEN RNA interference SNAI1 (SNAIL) transcription factor shRNA Cell Line Tumor Transcription Activator-Like Effector Nucleases Rhabdomyosarcoma CRISPR Humans genome editing lcsh:QH301-705.5 CRISPR/Cas9 Gene knockout Gene Editing Gene knockdown Transcription activator-like effector nuclease designer nucleases Base Sequence Cas9 General Medicine 030104 developmental biology HEK293 Cells lcsh:Biology (General) 030220 oncology & carcinogenesis Snail Family Transcription Factors CRISPR-Cas Systems rhabdomyosarcoma (RMS) |
| Zdroj: | Cells Volume 9 Issue 5 Cells, Vol 9, Iss 1095, p 1095 (2020) |
| ISSN: | 2073-4409 |
| DOI: | 10.3390/cells9051095 |
| Popis: | Genome editing (GE) tools and RNA interference technology enable the modulation of gene expression in cancer research. While GE mediated by clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 or transcription activator-like effector nucleases (TALEN) activity can be used to induce gene knockouts, shRNA interacts with the targeted transcript, resulting in gene knockdown. Here, we compare three different methods for SNAI1 knockout or knockdown in rhabdomyosarcoma (RMS) cells. RMS is the most common sarcoma in children and its development has been previously associated with SNAI1 transcription factor activity. To investigate the role of SNAI1 in RMS development, we compared CRISPR/Cas9, TALEN, and shRNA tools to identify the most efficient tool for the modulation of SNAI1 expression with biological effects. Subsequently, the genome sequence, transcript levels, and protein expression of SNAI1 were evaluated. The modulation of SNAI1 using three different approaches affected the morphology of the cells and modulated the expression of myogenic factors and HDAC1. Our study revealed a similar effectiveness of the tested methods. Nevertheless, the low efficiency of the GE tools was a limiting factor in obtaining biallelic gene knockouts. To conclude, we established and characterized three different models of SNAI1 knockout and knockdown that might be used in further studies investigating the role of SNAI1 in RMS. |
| Databáze: | OpenAIRE |
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