Killian, Tobias and Buntz, Annette and Herlet, Teresa and Seul, Heike and Mundigl, Olaf and Laengst, Gernot and Brinkmann, Ulrich (2019) Antibody-targeted chromatin enables effective intracellular delivery and functionality of CRISPR/Cas9 expression plasmids. NUCLEIC ACIDS RESEARCH, 47 (10). ISSN 0305-1048, 1362-4962
Full text not available from this repository. (Request a copy)Abstract
We report a novel system for efficient and specific targeted delivery of large nucleic acids to and into cells. Plasmid DNA and core histones were assembled to chromatin by salt gradient dialysis and subsequently connected to bispecific antibody derivatives (bsAbs) via a nucleic acid binding peptide bridge. The resulting reconstituted vehicles termed plasmid-chromatin' deliver packaged nucleic acids to and into cells expressing antigens that are recognized by the bsAb, enabling intracellular functionality without detectable cytotoxicity. High efficiency of intracellular nucleic acid delivery is revealed by intracellular expression of plasmid encoded genes in most (approximate to 90%) target cells to which the vehicles were applied under normal growth/medium conditions in nanomolar concentrations. Specific targeting, uptake and transgene expression depends on antibody-mediated cell surface binding: plasmid chromatin of identical composition but with non-targeting bsAbs or without bsAbs is ineffective. Examples that demonstrate applicability, specificity and efficacy of antibody-targeted plasmid chromatin include reporter gene constructs as well as plasmids that enable CRISPR/Cas9 mediated genome editing of target cells.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | VIRUS-LIKE PARTICLES; VIVO GENE-THERAPY; VIRAL VECTORS; DNA DELIVERY; NONVIRAL VECTORS; CELLS; PEPTIDE; PAYLOAD; HISTONES; PROGRESS; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Prof. Dr. Gernot Längst |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Apr 2020 05:00 |
| Last Modified: | 14 Apr 2020 05:00 |
| URI: | https://pred.uni-regensburg.de/id/eprint/26854 |
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