Engineered Plasmids for Fluorescent Tracking

Plasmids expressing foldon–GFP–PCP [a fusion protein consisting of the PP7 coat protein (PCP), green fluorescent protein (GFP) and a T4 fibritin trimeric motif foldon] were gifts from Chun-Qing Song (14 (link)). The single guide RNAs (sgRNAs) used in this study were constructed by modifying pX330-mitf (Addgene, 69801) or pLH-sgRNA1-2XPP7 (Addgene, 75390). The fragments of sgRNA were subcloned into the backbone vector using the NEB Gibson assembly kit (NEB, E2611L). All gRNAs in this research are listed in Supplementary Tables S1 and S2.

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Tian R., Huang Z., Li L., Yuan J., Zhang Q., Meng L., Lang B., Hong Y., Zhong C., Tian X., Cui Z., Jin Z., Liu J., Huang Z., Wang Y., Chen Y, & Hu Z. (2023). HPV integration generates a cellular super-enhancer which functions as ecDNA to regulate genome-wide transcription. Nucleic Acids Research, 51(9), 4237-4251.

Publication 2023

NEB Gibson assembly kit

Backbone Gifts Green fluorescent protein Mitf Plasmids Protein Single guide rnas Vector

Corresponding Organization : Zhongnan Hospital of Wuhan University

Other organizations : Renmin Hospital of Wuhan University, Huazhong University of Science and Technology, Central Hospital of Wuhan, Macao Polytechnic University, Peking University

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Variable analysis

independent variables

Plasmids expressing foldon–GFP–PCP
Single guide RNAs (sgRNAs) constructed by modifying pX330-mitf or pLH-sgRNA1-2XPP7

dependent variables

Unspecified

control variables

Unspecified

controls

Positive control: Unspecified
Negative control: Unspecified

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