Artificial Assemblies with Cooperative DNA Recognition by Zutao YU

Author:Zutao YU
Language: eng
Format: epub, pdf
ISBN: 9789811544231
Publisher: Springer Singapore

2.4 Cell Study of PIP–HoGu System

Therefore, because we had compellingly demonstrated its cooperativity in several in vitro assay systems, we were encouraged to apply PIP–HoGu to a cell-based assay. The estrogen response element (ERE) is the specific target motif of the estrogen receptor α (ERα) homodimer, which induces significant downstream gene activation (Fig. 2.7a) [44]. In ERα-positive, 17β-estradiol (E2)-stimulated T47D-KBluc cells that highly express luciferase after binding of three tandem ERα TF pairs, PIPs Ada3, Cyd2, and Ada3–Cyd2 that bound to the ERE consensus half-site (5′-WGGWCW-3′) were tested for 48 h together with the delivery reagent endoporter (Figs. 2.7b, c and 2.8) [45].

Fig. 2.7Cell-based assay of PIP–HoGu. a Crystal structure of estrogen receptor α (ERα) homodimer and DNA sequence. b The sequence of estrogen response element (ERE)-driven luciferase in T47D-KBLuc cells. c The structural design of Ada3 and Cyd2 targeting ERE sites (Fig. 2.8). d Conjugates Ada3, Cyd2, and Ada3–Cyd2 were screened for suppression of ERE-driven luciferase normalized to total protein using the BCA protein assay

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