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Title: | Análisis de variabilidad de secuencia in vivo de RNAs circulares con ribozimas |
Author: | Bañuls Tornero, Marc |
Tutor: | Maynou Fernández, Joan |
Others: | Maceira, Marc Canovas Izquierdo, Javier Luis |
Abstract: | Non-autonomous LTR retrotransposons that are expressed as circular RNAs with HHR ribozymes, named as retrozymes, have recently been discovered. In this study we carried out a detailed analysis of the natural variability of a Fragaria ananassa retrozyme under different conditions. Specifically, we studied the degree of in vivo variability in the chosen retrozyme, the number of variants that appear over time, whether the circularization of the RNA of the retrozyme affects or not the variability of the retrozyme, and the effect of the variability in its secondary structure. Different constructs were designed for the genetic transformation of two plant models to purify and extract the RNA and perform Sanger sequencing. The sequences obtained were ordered and used to perform an analysis of variants using inhouse scripts with Biopython, bwa-mem, BCFtools and SAMtools, while the secondary structures were obtained using the RNAalifold web service. The experiments carried out confirmed the presence of the same group of stable variants, which would indicate some type of adaptive advantage or specific edition of the circRNAs. Furthermore, it has been shown that the variability in mutated retrozymes that encode linear RNAs is minimal compared to the variability of circRNAs. The specific function of these stable variants remains to be determined in future experiments, since it does not seem to influence in conserved motifs or its secondary structure. |
Keywords: | circRNA retrozyme LTR retrotransposons |
Document type: | info:eu-repo/semantics/masterThesis |
Issue Date: | 24-Jun-2020 |
Publication license: | http://creativecommons.org/licenses/by-nc-nd/3.0/es/ |
Appears in Collections: | Trabajos finales de carrera, trabajos de investigación, etc. |
Files in This Item:
File | Description | Size | Format | |
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mbanulstTFM0620memoria.pdf | Memoria del TFM | 720,08 kB | Adobe PDF | View/Open |
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