Covalent coupling of high-affinity ligands to the surface of viral vector particles by protein trans-splicing mediates cell type-specific gene transfer

Muik, Alexander; Reul, Johanna; Friedel, Thorsten; Muth, Anke; Hartmann, Karen Patricia; Schneider, Irene C.; Münch, Robert C.; Buchholz, Christian J.

doi:10.1016/j.biomaterials.2017.07.032

Cited by 40 publications

(40 citation statements)

References 39 publications

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“…Split inteins have been used to generate a C-terminal thioester on a protein, with subsequent conjugation to flock house virus via native chemical ligation ( 54 ). Split inteins have also been used to functionalize adeno-associated virus particles with scFvs or DARPins, showing a coupling yield up to 10% ( 69 ). Some of the fastest split inteins operate via a reactive cysteine, which may be problematic with disulfide-containing antigens (see Classic Approaches to Modular VLNP Decoration ).…”

Section: Latest Approaches In Post-assembly Decoration Of Particlesmentioning

confidence: 99%

New Routes and Opportunities for Modular Construction of Particulate Vaccines: Stick, Click, and Glue

2018

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Vaccines based on virus-like particles (VLPs) can induce potent B cell responses. Some non-chimeric VLP-based vaccines are highly successful licensed products (e.g., hepatitis B surface antigen VLPs as a hepatitis B virus vaccine). Chimeric VLPs are designed to take advantage of the VLP framework by decorating the VLP with a different antigen. Despite decades of effort, there have been few licensed chimeric VLP vaccines. Classic approaches to create chimeric VLPs are either genetic fusion or chemical conjugation, using cross-linkers from lysine on the VLP to cysteine on the antigen. We describe the principles that make these classic approaches challenging, in particular for complex, full-length antigens bearing multiple post-translational modifications. We then review recent advances in conjugation approaches for protein-based non-enveloped VLPs or nanoparticles, to overcome such challenges. This includes the use of strong non-covalent assembly methods (stick), unnatural amino acids for bio-orthogonal chemistry (click), and spontaneous isopeptide bond formation by SpyTag/SpyCatcher (glue). Existing applications of these methods are outlined and we critically consider the key practical issues, with particular insight on Tag/Catcher plug-and-display decoration. Finally, we highlight the potential for modular particle decoration to accelerate vaccine generation and prepare for pandemic threats in human and veterinary realms.

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Section: Latest Approaches In Post-assembly Decoration Of Particlesmentioning

confidence: 99%

New Routes and Opportunities for Modular Construction of Particulate Vaccines: Stick, Click, and Glue

2018

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“…Efforts to improve this have made use of functionalized nanoparticles, stabilizing chemical modifications and cell-type specific ligand conjugates to improve efficacy, efficiency and specificity of delivery and reduce immunogenicity (79, 80). shRNAs can be delivered using viral vectors, such as recombinant adeno-associated viruses (rAAVs) programmed to have cell-type specific tropisms (81–83). These strategies are already being implemented in numerous clinical trials for the knockdown of disease causing mRNAs, highlighting the potential use of this technology for targeting lncRNAs (79).…”

Section: An Opportunity For Novel Immunotherapiesmentioning

confidence: 99%

The lncRNA Connection Between Cellular Metabolism and Epigenetics in Trained Immunity

et al. 2019

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Trained immunity describes the ability of innate immune cells to form immunological memories of prior encounters with pathogens. Recollection of these memories during a secondary encounter manifests a broadly enhanced inflammatory response characterized by the increased transcription of innate immune genes. Despite this phenomenon having been described over a decade ago, our understanding of the molecular mechanisms responsible for this phenotype is still incomplete. Here we present an overview of the molecular events that lead to training. For the first time, we highlight the mechanistic role of a novel class of long non-coding RNAs (lncRNAs) in the establishment and maintenance of discrete, long lasting epigenetic modifications that are causal to the trained immune response. This recent insight fills in significant gaps in our understanding of trained immunity and reveals novel ways to exploit trained immunity for therapeutic purposes.

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“…In this respect, we could recently demonstrate that ratios of VP1, VP2, and VP3 in assembled AAV capsids can vary with the polypeptide fused to VP2 and affect the overall yield. 35 Also for LV particles the DARPin can impact on the incorporation efficiency of the MV H fusion protein with a direct consequence for the overall amount of transduction-competent particles ( Figures S9 A and S9 B). 18 Interestingly, less than half of the selected GluA4-DARPins and about one-third of the tested CD105-DARPins mediated gene transfer of LV or AAV vectors.…”

Section: Discussionmentioning

confidence: 99%

A Library-Based Screening Strategy for the Identification of DARPins as Ligands for Receptor-Targeted AAV and Lentiviral Vectors

Hartmann

Münch

Freiling

et al. 2018

Molecular Therapy - Methods & Clinical Development

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Delivering genes selectively to the therapeutically relevant cell type is among the prime goals of vector development. Here, we present a high-throughput selection and screening process that identifies designed ankyrin repeat proteins (DARPins) optimally suited for receptor-targeted gene delivery using adeno-associated viral (AAV) and lentiviral (LV) vectors. In particular, the process includes expression, purification, and in situ biotinylation of the extracellular domains of target receptors as Fc fusion proteins in mammalian cells and the selection of high-affinity binders by ribosome display from DARPin libraries each covering more than 1012 variants. This way, DARPins specific for the glutamate receptor subunit GluA4, the endothelial surface marker CD105, and the natural killer cell marker NKp46 were generated. The identification of DARPins best suited for gene delivery was achieved by screening small-scale vector productions. Both LV and AAV particles displaying the selected DARPins transduced only cells expressing the corresponding target receptor. The data confirm that a straightforward process for the generation of receptor-targeted viral vectors has been established. Moreover, biochemical analysis of a panel of DARPins revealed that their functional cell-surface expression as fusion proteins is more relevant for efficient gene delivery by LV particles than functional binding affinity.

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Covalent coupling of high-affinity ligands to the surface of viral vector particles by protein trans-splicing mediates cell type-specific gene transfer

Cited by 40 publications

References 39 publications

New Routes and Opportunities for Modular Construction of Particulate Vaccines: Stick, Click, and Glue

New Routes and Opportunities for Modular Construction of Particulate Vaccines: Stick, Click, and Glue

The lncRNA Connection Between Cellular Metabolism and Epigenetics in Trained Immunity

A Library-Based Screening Strategy for the Identification of DARPins as Ligands for Receptor-Targeted AAV and Lentiviral Vectors

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