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| − | Peppers allow simple and robust imaging of diverse RNA species in live cells with minimal perturbation of the target RNA’s transcription, localization and translation.The map of the plasmid was listed below (Fig 1).
| + | Given the fact that lncRNA MALAT1 could act as miRNA sponge, we designed the sequence that could bind to miR-22 based on MALAT1 sequence. |
| − | Within pcDNA3-MCS-nPepper, the major components comprise the following modules: CMV, MCS, Pepper array, bGH polyA, AmpR promoter, AmpR, pUC ori, and SV40 ori. Both CMV and AmpR promoter function as promoters to initiate plasmid transcription. Following the CMV promoter, the MCS module can integrate the necessary nucleotide sequence, where NheI, HindIII, Acc651, and EcoRI can be employed as cutting enzymes for splicing. The Pepper array is transcribed simultaneously throughout the process and can be visualized using the fluorescent dye HBC for imaging purposes. The bGH polyA cap structure is employed to regulate the transcription process of the target gene. The AmpR promoter within the plasmid is utilized to initiate the transcription of ampicillin resistance(AmpR), facilitating the selection of successfully transformed bacteria on media.
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Revision as of 06:30, 23 September 2023
Given the fact that lncRNA MALAT1 could act as miRNA sponge, we designed the sequence that could bind to miR-22 based on MALAT1 sequence.
