Development of a novel method for rapid cloning of shRNA vectors, which successfully knocked down CD44 in mesenchymal triple-negative breast cancer cells

作     者：Lei Zhou Dandan Sheng Qiaodan Deng Dong Wang Suling Liu 

作者机构：The CAS Key Laboratory of Innate Immunity and Chronic DiseaseSchool of Life Sciences and Medical CenterUniversity of Science&Technology of ChinaHefei 230027AnhuiP.R.China Fudan University Shanghai Cancer Center&Institutes of Biomedical SciencesShanghai Medical CollegeKey Laboratory of Breast Cancer in ShanghaiInnovation Center for Cell Signaling NetworkCancer InstitutesDepartment of Breast SurgeryFudan UniversityShanghai 200032P.R.China 

出 版 物：《Cancer Communications》 (癌症通讯（英文）)

年 卷 期：2018年第38卷第1期

页      面：617-621页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：by the National Key Research and Development Program of China(Stem Cell and Translational Research 2016YFA0101202) National Nature Science Foundation of China Grants(81530075 and 81472741) Fudan University Research Foundation(IDH 1340042) Research Foundation of the Fudan University Shanghai Cancer Center(YJRC1603) the Ministry of Science and Technology of China Grant(2015CB553800) 

主　　题：rapid duplex companies 

摘      要：Dear Editor,Since the discovery of short hairpin RNA(shRNA)vec-tor-mediated RNA interference(RNAi),this technology has been widely used in cancer research for its specific-ity,potency,and ***,researchers may find it costly to purchase commercial vectors from bio-companies or time-and labor-consuming to construct their own shRNA vectors using traditional method by inserting annealed duplex into digested *** intensive efforts to accelerate shRNA vector cloning in laboratories,the development of a reliable,rapid,conven-ient,and cost-effective method is still in great *** this end,we developed a novel method named SuperSH(Super rapid cloning of shRNA vector)for the effective and rapid construction of shRNA-expressing vectors based on high-performance DNA polymerase and seamless cloning technique[1](Additional file 1:Fig-ure S1a;the detailed methods can be found in Additional file 1).In our SuperSH method,the shRNA sequences are introduced into the vector via a pair of polymerase chain reaction(PCR)primers rather than via annealed *** detail,the 3′ends of the primers are designed to bind the template to initiate a PCR to amplify the vector back-bone,and the 5′portions are designed to introduce the sequences of interest as well as to form a short homol-ogous arm for subsequent recombination via seamless cloning[1].