PUBLICATION
Embryo-scale reverse genetics at single-cell resolution
- Authors
- Saunders, L.M., Srivatsan, S.R., Duran, M., Dorrity, M.W., Ewing, B., Linbo, T.H., Shendure, J., Raible, D.W., Moens, C.B., Kimelman, D., Trapnell, C.
- ID
- ZDB-PUB-231116-5
- Date
- 2023
- Source
- Nature 623(7988): 782-791 (Journal)
- Registered Authors
- Kimelman, David, Linbo, Tor, Moens, Cecilia, Raible, David
- Keywords
- none
- Datasets
- GEO:GSE202639
- MeSH Terms
-
- Animals
- Embryo, Mammalian*/embryology
- Embryo, Mammalian*/metabolism
- Gene Expression Profiling
- Gene Expression Regulation, Developmental
- Mutation
- Notochord/cytology
- Notochord/embryology
- Reverse Genetics*/methods
- Single-Cell Analysis*/methods
- Transcriptome/genetics
- Zebrafish*/embryology
- Zebrafish*/genetics
- PubMed
- 37968389 Full text @ Nature
Citation
Saunders, L.M., Srivatsan, S.R., Duran, M., Dorrity, M.W., Ewing, B., Linbo, T.H., Shendure, J., Raible, D.W., Moens, C.B., Kimelman, D., Trapnell, C. (2023) Embryo-scale reverse genetics at single-cell resolution. Nature. 623(7988):782-791.
Abstract
The maturation of single-cell transcriptomic technologies has facilitated the generation of comprehensive cellular atlases from whole embryos1-4. A majority of these data, however, has been collected from wild-type embryos without an appreciation for the latent variation that is present in development. Here we present the 'zebrafish single-cell atlas of perturbed embryos': single-cell transcriptomic data from 1,812 individually resolved developing zebrafish embryos, encompassing 19 timepoints, 23 genetic perturbations and a total of 3.2 million cells. The high degree of replication in our study (eight or more embryos per condition) enables us to estimate the variance in cell type abundance organism-wide and to detect perturbation-dependent deviance in cell type composition relative to wild-type embryos. Our approach is sensitive to rare cell types, resolving developmental trajectories and genetic dependencies in the cranial ganglia neurons, a cell population that comprises less than 1% of the embryo. Additionally, time-series profiling of individual mutants identified a group of brachyury-independent cells with strikingly similar transcriptomes to notochord sheath cells, leading to new hypotheses about early origins of the skull. We anticipate that standardized collection of high-resolution, organism-scale single-cell data from large numbers of individual embryos will enable mapping of the genetic dependencies of zebrafish cell types, while also addressing longstanding challenges in developmental genetics, including the cellular and transcriptional plasticity underlying phenotypic diversity across individuals.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping