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Active Nuclear Import of Mammalian Cell-Expressible DNA Origami.

ABSTRACT: DNA origami enables the creation of complex 3D shapes from genetic material. Future uses could include the delivery of genetic instructions to cells, but nuclear import remains a major barrier to gene delivery due to the impermeability of the nuclear membrane. Here we realize active nuclear import of DNA origami objects in dividing and chemically arrested mammalian cells. We developed a custom DNA origami single-strand scaffold featuring a mammalian-cell expressible reporter gene (mCherry) and multiple Simian virus 40 (SV40) derived DNA nuclear targeting sequences (DTS). Inclusion of the DTS within DNA origami rescued gene expression in arrested cells, indicating that active transport into the nucleus occurs. Our work successfully adapts mechanisms known from viruses to promote the cellular expression of genetic instructions encoded within DNA origami objects.

SUBMITTER: Liedl A

PROVIDER: S-EPMC9999407 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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Active Nuclear Import of Mammalian Cell-Expressible DNA Origami.

Liedl Anna A Grießing Johanna J Kretzmann Jessica A JA Dietz Hendrik H

Journal of the American Chemical Society 20230224 9

DNA origami enables the creation of complex 3D shapes from genetic material. Future uses could include the delivery of genetic instructions to cells, but nuclear import remains a major barrier to gene delivery due to the impermeability of the nuclear membrane. Here we realize active nuclear import of DNA origami objects in dividing and chemically arrested mammalian cells. We developed a custom DNA origami single-strand scaffold featuring a mammalian-cell expressible reporter gene (mCherry) and m ...[more]

PMID: 36828394

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