PUBLICATION
Functionally characterizing obesity-susceptibility genes using CRISPR/Cas9, in vivo imaging and deep learning
- Authors
- Mazzaferro, E., Mujica, E., Zhang, H., Emmanouilidou, A., Jenseit, A., Evcimen, B., Metzendorf, C., Dethlefsen, O., Loos, R.J., Vienberg, S.G., Larsson, A., Allalou, A., den Hoed, M.
- ID
- ZDB-PUB-250214-22
- Date
- 2025
- Source
- Scientific Reports 15: 54085408 (Journal)
- Registered Authors
- Keywords
- CRISPR/Cas9, Deep learning, Fluorescence microscopy, Image analysis, Zebrafish
- MeSH Terms
-
- Genetic Predisposition to Disease
- Adipocytes/metabolism
- Animals
- Obesity*/genetics
- Obesity*/metabolism
- Lipid Metabolism/genetics
- Deep Learning*
- Mutation
- CRISPR-Cas Systems*
- Zebrafish*/genetics
- PubMed
- 39948378 Full text @ Sci. Rep.
Citation
Mazzaferro, E., Mujica, E., Zhang, H., Emmanouilidou, A., Jenseit, A., Evcimen, B., Metzendorf, C., Dethlefsen, O., Loos, R.J., Vienberg, S.G., Larsson, A., Allalou, A., den Hoed, M. (2025) Functionally characterizing obesity-susceptibility genes using CRISPR/Cas9, in vivo imaging and deep learning. Scientific Reports. 15:54085408.
Abstract
Hundreds of loci have been robustly associated with obesity-related traits, but functional characterization of candidate genes remains a bottleneck. Aiming to systematically characterize candidate genes for a role in accumulation of lipids in adipocytes and other cardiometabolic traits, we developed a pipeline using CRISPR/Cas9, non-invasive, semi-automated fluorescence imaging and deep learning-based image analysis in live zebrafish larvae. Results from a dietary intervention show that 5 days of overfeeding is sufficient to increase the odds of lipid accumulation in adipocytes by 10 days post-fertilization (dpf, n = 275). However, subsequent experiments show that across 12 to 16 established obesity genes, 10 dpf is too early to detect an effect of CRISPR/Cas9-induced mutations on lipid accumulation in adipocytes (n = 1014), and effects on food intake at 8 dpf (n = 1127) are inconsistent with earlier results from mammals. Despite this, we observe effects of CRISPR/Cas9-induced mutations on ectopic accumulation of lipids in the vasculature (sh2b1 and sim1b) and liver (bdnf); as well as on body size (pcsk1, pomca, irs1); whole-body LDLc and/or total cholesterol content (irs2b and sh2b1); and pancreatic beta cell traits and/or glucose content (pcsk1, pomca, and sim1a). Taken together, our results illustrate that CRISPR/Cas9- and image-based experiments in zebrafish larvae can highlight direct effects of obesity genes on cardiometabolic traits, unconfounded by their - not yet apparent - effect on excess adiposity.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping