Editing of Long Noncoding RNA DANCR Gene Locus Using Genetic Engineering Tools

Abstract

Long non-coding RNAs (lncRNAs) are defined as RNA transcripts that are longer than 200 nucleotides. By definition, these RNAs must not have open reading frames that encode proteins. We study the roles of lncRNA in DNA damage response (DDR). For this, we performed transcriptome analysis in human colorectal cancer cell line and mouse embryo fibroblast cells to elucidate differentially expressed lncRNAs in doxorubicin induced DNA damage. We identified DANCR lncRNA that is upregulated in DDR. The possible molecular functions of DANCR in DDR and in cell death remain to be discovered. In order to study functions of DANCR, we planned to create DANCR-/- cell lines using CRISPR/Cas9 genome engineering and editing tool. We aimed to remove DANCR from the genome without interfering the neighboring genes. After the cells are transfected with CRISPR/Cas9 constructs, they will be genotyped for confirming the deletion of the gene and they will be used to investigate the effects of the absence of DANCR in DDR with proliferation and apoptosis assays. This study will help to identify new players in DDR that will eventually shed light onto tumorigenesis and cancer treatment.