NEPA21: Organoid Applicaitons and Publications
A SONIDEL overview of NEPA21 use in organoid electroporation, including representative protocols, organoid result snapshots, and publication coverage across diverse human and murine organoid systems.
The NEPA21 is presented here from the perspective of optimising delivered energy for delicate organoid targets. The underlying workflow logic is to use only the energy needed to porate the membrane, helping researchers preserve viability while maintaining practical gene-delivery performance.
The NAPA21 platform supports both dissociated organoids via the CU540 cuvette electrode and whole organoids via the CUY650P1 electrode, alongside a broader electrode ecosystem intended to expand experimental flexibility.
Why organoid groups evaluate the NEPA21
For sensitive organoid workflows, the central practical question is not only whether cargo enters the target, but whether membrane opening, cargo transfer, and recovery can be tuned tightly enough to preserve biological usefulness downstream. This page positions the NEPA21 around that requirement: fine control over delivered energy, lower-current workflows relative to simpler electroporation approaches, and flexibility across dissociated and intact organoid formats.
The link to the full NEPA21 Organoid Publication List points to adoption in organoid laboratories, including the Organoid Group at the Hubrecht Institute, and to a broad publication base spanning intestinal, colon, brain, airway, breast, pancreatic, liver, ovarian, conjunctival, lacrimal, thymic, and other organoid systems.
Methodologically, the most relevant differentiators here are separate poring/transfer-style control logic, multiple electrode options, and workflow support for both cluster/single-cell style preparation and intact-organoid handling.
Representative protocol highlights
We highlights two representative protocol routes and several practical optimisation notes for gastrointestinal and intestinal organoid workflows.
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Multiple gene knockout in mouse small intestinal organoids
A cited step-by-step JoVE protocol focused on CRISPR-concatemer-based knockout in mouse small intestinal organoids.
Human intestinal and GI organoid engineering
We highlight both Efficient genetic engineering of human intestinal organoids using electroporation and the JoVE protocol Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids.
Workflow details called out in publications
Clusters of roughly 10–15 cells were used instead of full single-cell dissociation, plasmids of 4.2 kb and 9.3 kb were compared, Cas9 plasmid was also tested, the workflow was described as taking 1 day instead of 4 days, and a 40-minute regeneration step after electroporation was highlighted as beneficial for survivability and especially large-plasmid transfection efficiency.
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Client Reported transfection examples
Client laboratory verified results snapshot
Additional know-how data are available from SONIDEL on request.
| Cell / organoid type | Viability | Transfection efficiency |
|---|---|---|
| Human Normal Fundic Gastric Organoids | 68% | — |
| Mouse Fundic Gastric Organoids | 65% | — |
| Mouse Colorectal Cancer Organoids | 100% | 50% |
Colon organoid example from Keio University
The PDF includes a TP53 knockout colon organoid example in which TP53 KO organoids were selected with Nutlin-3. The accompanying note states that TP53 KO organoids tolerated Nutlin-3, whereas WT colon organoids did not grow under Nutlin-3-supplemented conditions.
Publication coverage across organoid systems
The NEPA21 publication footprint spans a wide set of human and murine organoid contexts. Note the link to a full publication listing: [NEPA21 Organoid Publications]
Selected publication highlights
Some representative publications.
Foundational intestinal organoid engineering
Efficient genetic engineering of human intestinal organoids using electroporation
Nat Protoc. 2015
Modeling colorectal cancer using CRISPR-Cas9-mediated engineering of human intestinal organoids
Nat Med. 2015
Differentiation and CRISPR-Cas9-mediated genetic engineering of human intestinal organoids
STAR Protoc. 2022
GI organoid protocol and CRC examples
Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids
J Vis Exp. 2020
A protocol for efficient CRISPR-Cas9-mediated knock-in in colorectal cancer patient-derived organoids
STAR Protoc. 2021
Recapitulating the adenoma–carcinoma sequence in mismatch-repair-deficient human colon organoids
2024
Brain and neuronal organoid examples
Human fetal brain self-organizes into long-term expanding organoids
Cell. 2024
Calcium-Enhanced Medium-Based Delivery of Splice Modulating Antisense Oligonucleotides in hiPSC-derived neuronal models
Biomedicines. 2024
Contribution of rare coding variants to microcephaly in human forebrain organoid-linked systems
Genome Med. 2025
Airway and ocular surface models
Modelling of primary ciliary dyskinesia using patient-derived airway organoids
EMBO Rep. 2021
Drug Repurposing for Cystic Fibrosis in Nasal Organoids
Int J Mol Sci. 2022
Human conjunctiva organoids to study ocular surface homeostasis and disease
Cell Stem Cell. 2024
Pancreatic and liver-linked examples
USP25 promotes pathological HIF-1-driven metabolic reprogramming in pancreatic cancer
Nat Commun. 2022
Engineered human hepatocyte organoids enable CRISPR-based target discovery and drug screening for steatosis
Nat Biotechnol. 2023
Establishment of human fetal hepatocyte organoids and CRISPR-Cas9 knockin/knockout
Nat Protoc. 2020
Breast and ovarian organoid examples
Long-term culture, genetic manipulation and xenotransplantation of human normal and breast cancer organoids
Nat Protoc. 2021
A Living Biobank of Breast Cancer Organoids Captures Disease Heterogeneity
Cell. 2018
Organoid platinum-resistance model identifies KRT17 as a biomarker in ovarian cancer
iScience. 2025
Murine intestinal and glandular protocols
A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
J Vis Exp. 2017
Establishment, Maintenance, Differentiation, Genetic Manipulation, and Transplantation of Mouse and Human Lacrimal Gland Organoids
J Vis Exp. 2023
Derivation of functional thymic epithelial organoid lines from adult murine thymus
Cell Rep. 2024
Further systems shown in the PDF
Fallopian tube epithelium, endometrial organoids, head and neck cancer organoids, human gastric assembloids, biopsy-derived duodenal organoids, murine normal colorectal models, and organoid-derived neural progenitor cell workflows are all represented in the attached bibliography.
What this publication base signals
Taken together, the full publication list positions the NEPA21 as a practical option for early-stage engineering, perturbation, reporter delivery, and disease-modelling workflows across diverse organoid systems where viability, timing, and assay-readiness matter.
Summary
This page condenses why NEPA21 is used in organoid electroporation, what protocol features are emphasised, what result snapshots are shown, and how broad the publication base has become across human and murine organoid systems.
Discuss your organoid workflow with SONIDEL
Share the organoid type, cargo format, and whether you are working with dissociated organoids, intact organoids, or a gastrointestinal / translational model, and Sonidel can advise on an appropriate starting route.