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Upmeier zu Belzen, J.

Publications and source records attributed to Upmeier zu Belzen, J..

3 recordsLinked to original sources

Biologically Guided Variational Inference for Interpretable Multimodal Single-Cell Integration and Mechanistic Discovery

Multi-omics technologies allow detailed characterization of cell types and states across omics layers as well as chemical and genetic perturbations. Variational autoencoders have become a cornerstone of single-cell data integration; however, they are often implemented as black-box models, requiring post hoc interpretation using known markers, pathways, or regulators to make sense of their latent representations. NetworkVI fundamentally flips this paradigm by incorporating biological knowledge directly into the model architecture. By embedding co-regulation networks derived from topologically associated domains and structured ontologies such as the Gene Ontology (GO), NetworkVI introduces a biologically-informed inductive bias that promotes the preservation of meaningful variation during integration while enforcing interpretability at both the gene and GO levels. NetworkVI achieves state-of-the-art data integration, modality imputation, and cell label transfer across bimodal and trimodal datasets. Beyond integration, here we show that NetworkVI facilitates ontology-guided hypothesis generation by exploiting established associations between genes, structured cellular programs, and regulatory domains to interpretably model cellular identities. Furthermore, decomposition of GO activation spaces resolves lineage-specific functional states within immune cell types, including quiescent, inflammatory, and transitional monocyte subpopulations, that are invisible to transcriptomic clustering. NetworkVI prioritizes GO-term programs associated with immunosenescence, consistent with age-associated immune dysregulation and reveals candidate immune evasion mechanisms consistent with CD58 loss in a Perturb-CITE-seq melanoma dataset.

bioinformatics↗

Optogenetic control of Neisseria meningitidis Cas9 genome editing using an engineered, light-switchable anti-CRISPR protein

Optogenetic control of CRISPR-Cas9 systems has significantly improved our ability to perform genome perturbations in living cells with high precision in time and space. As new Cas orthologues with advantageous properties are rapidly being discovered and engineered, the need for straightforward strategies to control their activity via exogenous stimuli persists. The Cas9 from Neisseria meningitidis (Nme) is a particularly small and target-specific Cas9 orthologue, and thus of high interest for in vivo genome editing applications. Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein. Building on our previous Acr engineering work, we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa. Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation. Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface. Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.

synthetic biology↗

Computational design of anti-CRISPR proteins with improved inhibition potency and expanded specificity

Anti-CRISPR (Acr) proteins are bacteriophage-derived antagonists of CRISPR-Cas systems. To date, Acrs were obtained either by mining sequence databanks or experimentally screening phage collections, both of which yield a limited repertoire of naturally occurring variants. Here, we applied structure-based engineering on AcrIIC1, a broad-spectrum inhibitor of type II-C CRISPR systems, to improve its efficacy and expand its specificity. We first show that fusing exogenous protein domains into AcrIIC1 dramatically enhances inhibition of the natural Neisseria meningitidis Cas9 target. Then, using structure-guided design, we converted AcrIIC1 into AcrX, a potent inhibitor of the type II-A CRISPR-Cas9 from Staphylococcus aureus widely applied for in vivo genome editing. Our work introduces designer Acrs as important biotechnological tools and provides an innovative strategy to safeguard the CRISPR technology.

synthetic biology↗