Selected article for: "direct action and indirect action"
Author: K. Reddisiva Prasanth; Minato Hirano; W. Samuel Fagg; Eileen T. McAnarney; Chao Shan; Xuping Xie; Adam Hage; Colette A. Pietzsch; Alexander Bukreyev; Ricardo Rajsbaum; Pei-Yong Shi; Mark T. Bedford; Shelton S. Bradrick; Vineet Menachery; Mariano A. Garcia-Blanco
Title: Topoisomerase III-ß is required for efficient replication of positive-sense RNA viruses
  • Document date: 2020_3_27
    • ID: a0xfb9l4_5
    Snippet: The genetic approaches we utilized above do not distinguish between direct and indirect modes of action. To address whether or not TOP3B directly interacted with DENV-2 genomes we used UV crosslinking followed by RNA immunoprecipitation (CLIP). We carried out CLIP assays using a HEK-293T cell line that expressed a FLAG-tagged TOP3B upon doxycycline treatment and anti-FLAG antibodies to carry out the immunoprecipitation. FLAG-TOP3B preferentially .....
    Document: The genetic approaches we utilized above do not distinguish between direct and indirect modes of action. To address whether or not TOP3B directly interacted with DENV-2 genomes we used UV crosslinking followed by RNA immunoprecipitation (CLIP). We carried out CLIP assays using a HEK-293T cell line that expressed a FLAG-tagged TOP3B upon doxycycline treatment and anti-FLAG antibodies to carry out the immunoprecipitation. FLAG-TOP3B preferentially crosslinked to CELSR2 RNA, which was previously known to bind this topoisomerase 6 , relative to EEF1A1 RNA, which we use as a negative control ( Fig 1H) .

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