CSAG1 Knock out in clone E3 from colon cancer CC14 primary culture

In order to explore the role of CSAG1 in the metastatic potential of the highly metastatic clone E3 we knocked it out using CRISPR/Cas9 system . Here we aimed to explore the transcriptome of E3 cells on which the expression of CSAG1 has been supressed. Overall design: CSAG1 KO mutations were induced using CRISPR/Cas9 system in E3 clonal cells from primary colon cancer cells (CC14). These cells were cloned and mutations analysed by sequencing. One out of frame mutation with ATG deletion was chosen and functionnaly tested. CSAG1 KO cells were thenharvested and total RNA extracted using TRIzol Reagent (Ambion) following the manufacturer's protocol. Each condition was independently processed in triplicates and the libraries were prepared using Illumina TruSeq protocols. The samples were sequenced on a HiSeq Illumina 4000 machine with 50bp reads followed by standard analyses.