Part:BBa_K2908000

- We identified genes that were differentially expressed in human triple negative breast cancer (TNBC) cells compared to healthy tissues from the publicly available The Cancer Genome Atlas (TCGA) database [1] using R (version 3.0.2). We followed standard procedures to calculate differentially expressed genes between cancer cases compared to controls using t test. The p values were corrected for multiple testing using the Bonferroni correction. We then selected genes that were previously identified as transcription factors (TFs) from the literature. From these genes (ESR1, FOXA1, GATA3, E2F1, PTTG1, UHRF1, FOXM1, MYBL2, PITX1), we selected the ones that verified as the most specific in MDA-MB-231 cells (TNBC cell line) compared to HBL-100 cells (normal breast cell line). (Refer to the Project/Demonstrate page in our wiki to see the results. Address:https://2019.igem.org/Team:CSU_CHINA/Demonstrate)
- Construction: We use the JASPER to get the TF binding sites (TF-BSs), and add different spacers between the tandem repeats of TF-BSs to avoid creating new binding sites in order to ensure the specificity.

This figure shows the results we browse using JASPAR by input homo sapiens GATA3.
Then we add a strong and short promoter named lateADEp to ensure the strength of the whole promoter.