Part:BBa_K2889002:Experience

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Applications of BBa_K2889002

1.cell migration

Cell migration is a central process in the development and maintenance of tumor. We cloned the full length of IL7-AS-S1 into PCDNA3.1 and transfected the plasmid to 786-O cells.Through in vitro scratch wound healing assay, overexpression of IL7-AS-S1 promoted cell migration of 786-O cells .These results suggested that IL7-AS-S1 contain key structural domains.(Fig. 1 and 2)

Overexpression of IL7-AS-S1 promotes 786-O cells migration. 786-O cells were transfected with pcDNA3.1-IL7-AS-S1 for 24 h. The cells growth areas were calculated with Image J at the time points of 0, 6, 12 and 24 h. Bar=100 μm. Data represents means ± SE from 3 independent experiments. *p<0.05 versus non-transfected cells.

2.IL7-AS contains a complicated secondary structure

In silico prediction of lncRNA secondary structure is another useful method to assign putative functions to non-coding transcripts, based upon the widely held assumption that highly folded structures impart functionality through binding interactions with 

proteins/nucleotides.Characterization of IL7-AS, IL7-AS-S1 and IL7-AS-S2 using RNAfold minimum free energy estimations predicted a highly folded secondary structure with several hairpin loops, which imply interact with protein (Fig. 3).

The optimal secondary structure with a minimum free energy. The secondary structures of IL7-AS-S1 were predicted by the RNAfold webserver (http://rna.tbi.univie.ac.at/cgi-bin/RNAfold.cgi). The structure is colored according to base-pairing probabilities. For unpaired regions, the color denotes the probability of being unpaired.

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