Part:BBa_K2889000:Experience

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K2889000

1.cell migration

1.1 Cell migration is a central process in the development and maintenance of tumor. We cloned the full length of IL7-AS-S2 into PCDNA3.1 and transfected the plasmid to 786-O cells.Through in vitro scratch wound healing assay, overexpression of IL7-AS-S2 promoted cell migration of 786-O cells.These results suggested that IL7-AS-S2 contain key structural domains(Fig. 1 and 2).

Overexpression of IL7-AS-S2 promotes 786-O cells migration. 786-O cells were transfected with pcDNA3.1-IL7-AS-S2 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.

1.2 In order to investigate whether cell migration induced by IL7-AS-S2 depends on the amount of IL7-AS-S2 transfection. We transfected the different concentration of IL7-AS-S2 into 786-0 cells.(Fig. 3 and 4)

2.IL7-AS-S2 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-S2 using RNAfold minimum free energy estimations predicted a highly folded secondary structure with several hairpin loops, which imply interact with protein (Fig. 5).

The optimal secondary structure with a minimum free energy. The secondary structures of IL7-AS-S2 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.

3.Bioinformatic prediction of IL7-AS-S2 function

LncRNAs perform the function through interacting with proteins. Considering IL7-AS-S2 can influence the cell migration, we want to predict proteins which can interact with IL7-AS-S2. Using the catRAPID omics algorithm (http://service.tartaglialab.com/page/catrapid_omics_group), we determined whether IL7-AS-S2 interacts with proteins. Results showed that the related RNA-binding proteins were involved in RNA-splicing/processing (CWC15, RU2A), and protein synthesis (EIF3G, SBDS) (table 1). Based on the proteins predicted, we estimated that IL7-AS-S2 may regulate RNA splicing or protein synthesis of some tumor related genes though interacting with several RNA-splicing or protein synthesis associated proteins.

User Reviews

UNIQ996379f57b055163-partinfo-00000000-QINU UNIQ996379f57b055163-partinfo-00000001-QINU