Proteomic and Transcriptome Profiling of G-Quadruplex Aptamers Developed for Cell Internalization

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

2 Scopus Citations
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Author(s)

  • Yang Zhang
  • Yang Wu
  • Hongjin Zheng
  • Hui Xi
  • Taoyu Ye
  • Chun-Yin Chan

Detail(s)

Original languageEnglish
Pages (from-to)5744-5753
Journal / PublicationAnalytical Chemistry
Volume93
Issue number14
Online published30 Mar 2021
Publication statusPublished - 13 Apr 2021

Abstract

Nucleic acid medicine is expected to be among the most promising next-generation therapies. Applications of nucleic acid in vivo are still challenging as a result of the difficulties in direct cell penetration without external assistance. To facilitate the cellular delivery of therapeutic nucleic acid, we developed cell-penetrating aptamers using cell-internalization Systematic Evolution of Ligands by EXponential enrichment (SELEX). Moreover, C20-4 min, a G-quadruplex-forming DNA aptamer, was discovered, showing a higher cell-penetrating capacity compared with other candidates, including AS1411. To verify the formation and understand the G-quadruplex folding topologies of enriched aptamer motifs, characteristic circular dichroism (CD) spectral features are analyzed. The CD spectra of C20-4 min strongly support the formation of parallel G-quadruplexes. Systematic analyses of the G-quadruplex regulation pathway have been performed by combining aptamer pull-down with mass spectrometry. We profiled G-quadruplex aptamers interacting with cellular proteins during internalization and identified helicases and GTPase proteins as cellular interacting partners. In addition, whole transcriptome analysis was performed to study the effects of G-quadruplex aptamers, revealing differentially expressed genes involved in the regulation of GTPase functions. Integrative analyses of transcriptome and proteomic have aided in understanding the functional hierarchy of molecular players in G-quadruplex nucleic acid mechanisms of internalization, which might facilitate developing a novel delivery system.

Citation Format(s)

Proteomic and Transcriptome Profiling of G-Quadruplex Aptamers Developed for Cell Internalization. / Zhang, Yang; Wu, Yang; Zheng, Hongjin; Xi, Hui; Ye, Taoyu; Chan, Chun-Yin; Kwok, Chun Kit.

In: Analytical Chemistry, Vol. 93, No. 14, 13.04.2021, p. 5744-5753.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review