Platforms for delivery of macromolecules to sites of DNA double-strand break repair

Zhen Cao, Deepika Goyal, Steffen E. Meiler, Yunfeng Zhou, William S. Dynan

Research output: Contribution to journalArticle

Abstract

Double-strand break (DSB) repair foci are important therapeutic targets. Here we describe platforms for delivery of macromolecules, nanomaterials and nanomedicines to repair foci. The strategy is based on the high affinity of the human 53BP1 protein for modified chromatin present at sites of DNA damage. As proof of concept, we created, expressed, and purified an engineered fragment of 53BP1 and coupled it to fluorescent streptavidin, a model cargo with no intrinsic affinity for repair foci. This binary complex was in turn coupled to the iron carrier protein, transferrin, which engages a high-affinity cell surface receptor. In a different version of the complex, transferrin was omitted and a protein transduction domain was incorporated directly into the primary structure of the 53BP1. These complexes were efficiently taken up into human osteosarcoma cells and synchronously released from endocytic vesicles by brief exposure to far-red light in the presence of the photosensitizer, disulfonated aluminum phthalocyanine. Upon release, the streptavidin cargo entered the nucleus and was recruited to repair foci. 53BP1-based platforms provide a method for targeted, temporally controlled delivery of macromolecular agents to sites of double-strand break repair. With the delivery platforms, we are capable to visualize, modify and redirect DSB repair pathways by coupling various nanomaterials to study machinery or manipulate for therapy purpose in the future.

Original languageEnglish (US)
Pages (from-to)2196-2204
Number of pages9
JournalArtificial Cells, Nanomedicine and Biotechnology
Volume47
Issue number1
DOIs
StatePublished - Dec 4 2019

Fingerprint

Streptavidin
Double-Stranded DNA Breaks
Nanostructures
Transferrin
Macromolecules
DNA
Repair
Nanomedicine
Transport Vesicles
Photosensitizing Agents
Cell Surface Receptors
Osteosarcoma
DNA Damage
Chromatin
Carrier Proteins
Iron
Light
Nanostructured materials
Therapeutics
Proteins

Keywords

  • 53BP1
  • Double strand-break repair
  • delivery
  • endocytosis
  • transferrin

ASJC Scopus subject areas

  • Biotechnology
  • Medicine (miscellaneous)
  • Biomedical Engineering
  • Pharmaceutical Science

Cite this

Platforms for delivery of macromolecules to sites of DNA double-strand break repair. / Cao, Zhen; Goyal, Deepika; Meiler, Steffen E.; Zhou, Yunfeng; Dynan, William S.

In: Artificial Cells, Nanomedicine and Biotechnology, Vol. 47, No. 1, 04.12.2019, p. 2196-2204.

Research output: Contribution to journalArticle

Cao, Zhen ; Goyal, Deepika ; Meiler, Steffen E. ; Zhou, Yunfeng ; Dynan, William S. / Platforms for delivery of macromolecules to sites of DNA double-strand break repair. In: Artificial Cells, Nanomedicine and Biotechnology. 2019 ; Vol. 47, No. 1. pp. 2196-2204.
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