PUBLICATION
Stage- and tissue-specific gene editing using 4-OHT-inducible Cas9 in whole organism
- Authors
- Li, Y., Zhang, W., Wei, Z., Li, H., Liu, X., Zheng, T., Aziz, T., Xing, C., Meng, A., Wu, X.
- ID
- ZDB-PUB-260103-1
- Date
- 2026
- Source
- The Journal of cell biology 225: (Journal)
- Registered Authors
- Meng, Anming, Xing, Cencan
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- CRISPR-Associated Protein 9*/genetics
- CRISPR-Associated Protein 9*/metabolism
- CRISPR-Cas Systems*/genetics
- Female
- Gene Editing*/methods
- Germ Cells/metabolism
- Humans
- Mice
- Organ Specificity
- Receptors, Estrogen/genetics
- Receptors, Estrogen/metabolism
- Tamoxifen*/analogs & derivatives
- Tamoxifen*/pharmacology
- Zebrafish*/embryology
- Zebrafish*/genetics
- PubMed
- 41481232 Full text @ J. Cell Biol.
Citation
Li, Y., Zhang, W., Wei, Z., Li, H., Liu, X., Zheng, T., Aziz, T., Xing, C., Meng, A., Wu, X. (2026) Stage- and tissue-specific gene editing using 4-OHT-inducible Cas9 in whole organism. The Journal of cell biology. 225:.
Abstract
Vertebrate genes function in specific tissues and stages, so their functional studies require conditional knockout or editing. In zebrafish, spatiotemporally inducible genome editing, particularly during early embryogenesis, remains challenging. Here, we establish inducible Cas9-based editing in defined cell types and stages. The nCas9ERT2 fusion protein, consisting of Cas9 and an estrogen receptor flanked by two nuclear localization signals, is usually located in the cytoplasm and efficiently translocated into nuclei upon 4-hydroxytamoxifen (4-OHT) treatment in cultured cells or embryos. As a proof of concept, we demonstrate that genes in primordial germ cells in embryos and germ cells in adult ovaries from a transgenic line with stable expression of nCas9ERT2 and gRNAs can be mutated by 4-OHT induction. The system also works in early mouse embryos. Thus, this inducible nCas9ERT2 approach enables temporospatial gene editing at the organismal level, expanding the tissue- and stage-specific gene-editing toolkit.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping