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Interfering XIST expression by CRISPR/Cas9 and TALEN
La Ting1, Ma Jianyang1, Shen Nan1,2, Tang Yuanjia1,2*
1Laboratory of Molecular Rheumatology of Institute of Health Sciences,Shanghai Jiaotong University School of Medicine (SJTUSM)&Shanghai Institutes for Biological Science (SIBS), Chinese Academy of Sciences (CAS), Shanghai 200025, China;
2Shanghai Institute of Rheumatology, Shanghai Jiaotong University School of Medicine Affiliated Renji Hospital, Shanghai 200001, China
Abstract: XIST is one of the long noncoding genes which could help to keep X chromosome silenced in mammals. The abnormity of X chromosome in activation increases the expression of X-linked genes， which is thought to represent a key event in oncogenesis and the occurrence of other diseases. Interfering XIST expression is necessary to study functions of XIST and associated diseases. In this study， we edited the known core promoter of XIST by CRISPR/Cas9 and TALEN in 293T cells. T7E1 assay and sequencing were used to detect the efficiency of mutation. Limiting dilution， fragment analysis and TA cloning were used to get the monoclonal cell lines expressing low XIST and identify their genotypes. Results showed that compared with the control group， both CRISPR/Cas9 and TALEN could mutate the core promoter and knock down the expression of XIST. In conclusion， the edited core promoter of XIST could knock down its expression. The results may contribute to new strategy about silencing long noncoding genes.
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