Team:Worldshaper-Wuhan/Design

Design

Design
References

Design

miRNA Sponges contain complementary binding sites to a miRNA of interest, which inhibit miRNA activity. This mechanism gives rise to our idea about fusing a sponge RNA based on the sequences of lncRNA with binding sites complementary to the sequence of miRNA to a plasmid that has reporter gene, EGFP-C1 for instance, which will monitor the expression of miRNA in the cells.

According to our previous study, we choose XIST to design our microRNA sensor. We downloaded the sequence of XIST from NCBI [1]and predicted miRNAs which could bind to XIST from TargetScan. We found miR-155 could bind to the sequence of XIST (Fig.2). miR-155 is an oncomiR that is overexpressed in several cancers, including breast cancer. A growing number of studies highlights the role of miR-155 in breast cancer drug resistance development. Given the fact that lncRNA could act as miRNA sponge, we designed the sequence that could bind to miR-155 based on XIST sequence. To improve the binding efficiency, miR-155 sensors contain three binding sites(Fig.3).

Fig.2. The pairing between miR-155 and XIST with binding sites is diagrammed. The miRNA seed region is showed by red.

Fig.3. The diagram of miR-155 sensor (pEGFP-miR-155-sponge-1)

Reference:

[1] NR_001564, https://www.ncbi.nlm.nih.gov/nuccore/NR_001564.2