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Image Search Results
Journal: Nucleic Acids Research
Article Title: Efficient circularization of protein-encoding RNAs via a novel cis -splicing system
doi: 10.1093/nar/gkae711
Figure Lengend Snippet: Impact of P9.0 structural design variants on circularization efficiency and RNA splicing. (A–F): (A, D) the schematic diagram of the circularization element using Tetrahymena as a model. ( A ) The molecular recognizer cannot facilitate efficient circularization without a paired structure (the design is referred to as Loop-ωG in this context). ( D ) Reintroducing paired structures (Arms) in R1 and R2 can reinstate circularization (the design is called Arm-Loop-ωG in this context). The BSS is within the open reading frame (ORF). ( B ) and ( E ) show the migration mode of the splicing products in the 2% E-Gel EX system. The lines indicate the product types of the corresponding bands, respectively. RNase R digests precursor RNAs but not circular RNAs, allowing circular RNAs to be enriched. The product purity of FA analysis is presented in percentage form. ( C ) and ( F ) show the fluorescence image of GFP expression in Huh7 cells transfected with circularized samples (before and after RNase R treatment). Scale bar: 50 μm. ( G ) Schematically illustrates the process of conducting RNA-seq analysis directly on circularized samples following column purification. After a two-step transesterification reaction, the results depict the splicing region of the GOI, encompassing the BSS, detected mutation types (deletion, insertion), and the corresponding probabilities of their occurrences, presented in percentages. InDel refers to insertion-deletion events. ( H ) Illustrates the circularization element using Tetrahymena as a model. In this depiction, P9.0 is substituted with a homologous arm adjacent to the ωG residue (the design is called Arm-ωG in this context). ( I ) Shows the migration mode of the splicing products in the 2% E-Gel EX system. The lines indicate the product types of the corresponding bands, respectively. RNase R digests precursor RNAs but not circular RNAs, allowing circular RNAs to be enriched. The product purity of FA analysis is presented in percentage form. ( J ) Displays the RNA-seq results showcasing the connecting region of the gene of interest (GOI) after splicing. This visualization includes the detected mutation types at the circularization site, such as insertion-deletion events (InDel).
Article Snippet: The sequences of
Techniques: Migration, Fluorescence, Expressing, Transfection, RNA Sequencing, Purification, Mutagenesis, Residue