ArticleNature cell biology2020
Fast and efficient generation of knock-in human organoids using homology-independent CRISPR-Cas9 precision genome editing.
Article in Nature cell biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 181 papers, 1 of them a synthesis that pooled it.
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Who cites it
181 citing papers in PubMed, 1 synthesis or guideline pooled it, 277 citations in OpenAlex.
- Application and prospect of organoid technology in breast cancer.Frontiers in immunology · 2024Pooled it
- Human Small Intestinal Organoids Differentiation Protocols Towards All Mature Epithelial Lineages.Methods in molecular biology (Clifton, N.J.) · 2027Article
- Tutorial: interrogating biology using organoid technologies.Nature protocols · 2026Review
- Quantitative imaging of dynamic processes in intestinal organoids.Nature protocols · 2026Review
- The Osteosarcoma Tumor Microenvironment: From Cellular Interactions to Advanced Preclinical Models.Cancers · 2026Review
- Liver Organoids: From Disease Modelling to Regenerative Medicine.Cell proliferation · 2026Review
- Patient-Derived Organoids in Gastrointestinal Disease: Current Applications, Limitations, and Future Perspectives.International journal of molecular sciences · 2026Review
- Harnessing human tumor organoids for cancer modeling and precision therapy.Protein & cell · 2026Review
- Patient-Derived Organoid-Based CRISPR Screens in Cancer Research: Applications, Advances, and Challenges.Cancer medicine · 2026Review
- The immediate cellular response to whole-genome doubling is conserved across polyploid contexts.bioRxiv : the preprint server for biology · 2026Article
- Synthetic developmental engineering of human liver organogenesis.Development (Cambridge, England) · 2026Review
- Brain organoids and genome editing: A new era in understanding human brain development and disorders.Neural regeneration research · 2026Article
- Patient-derived organoids in gynaecological cancers: emerging tools for therapy and disease modelling.Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico · 2026Review
- Stem cell-based tactics in the remodeling and treatment of intestinal diseases.Stem cell research & therapy · 2026Review
- Lessons on multicellular two- (2D) and three-dimensional (3D) culture in parasitology: Insights, challenges and future directions.International journal for parasitology · 2026Review
- A ZEB1-Neon knock-in uncovers traceable dynamics of epithelial-mesenchymal transition in tumors in vivo.BMC biology · 2026Article
- Article
- Progress and challenges in the development of advanced pancreatic cancer organoids.Journal of experimental & clinical cancer research : CR · 2026Review
- Translating Pharmacology from Animal Models to Organoids: Advancing Predictive Human Biology in Drug Discovery.Stem cell reviews and reports · 2026Review
- Monitoring biological effects of somatic cell genome editing.Nature reviews. Genetics · 2026Review
121 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
10 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR-Cas9 technology has revolutionized genome editing and is applicable to the organoid field. However, precise integration of exogenous DNA sequences into human organoids is lacking robust knock-in approaches. Here, we describe CRISPR-Cas9-mediated homology-independent organoid transgenesis (CRISPR-HOT), which enables efficient generation of knock-in human organoids representing different tissues. CRISPR-HOT avoids extensive cloning and outperforms homology directed repair (HDR) in achieving precise integration of exogenous DNA sequences into desired loci, without the necessity to inactivate TP53 in untransformed cells, which was previously used to increase HDR-mediated knock-in. CRISPR-HOT was used to fluorescently tag and visualize subcellular structural molecules and to generate reporter lines for rare intestinal cell types. A double reporter-in which the mitotic spindle was labelled by endogenously tagged tubulin and the cell membrane by endogenously tagged E-cadherin-uncovered modes of human hepatocyte division. Combining tubulin tagging with TP53 knock-out revealed that TP53 is involved in controlling hepatocyte ploidy and mitotic spindle fidelity. CRISPR-HOT simplifies genome editing in human organoids.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.