Omics · study · 2026
Highly efficient and low-mosaicism piggyBac transgenesis platform for rapid founder phenotyping [single-cell RNA-seq]
Listed in NCBI GEO
Pronuclear microinjection is the most widely used method for producing transgenic (Tg) animals.
Description
Because its efficiency is typically <20%, phenotypic characterization of Tg animals is usually performed from the next generation (F1) onwards. Conventional Cre-loxP-based conditional knockout (cKO) mouse production requires generating two genetically modified strains and multiple rounds of breeding before cKO mice are available for analysis.
Here, we optimized a piggyBac transposon-based method of Tg mouse production and established conditions under which nearly all founder (F0) embryos are transgenic. Using a single-cell RNA sequencing-based strategy, we characterized mosaicism in F0 embryos and demonstrated that piggyBac-mediated transgene integration occurs early in embryonic development. We also achieved ~70% efficiency in generating bacterial-artificial-chromosome-Tg mice.
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By combining this method with genome editing, we developed a strategy for tissue-specific-knockout phenotyping in the F0 generation. Our findings establish a highly efficient piggyBac-based platform that expands experimental options for Tg animal production by supporting rapid F0-based phenotypic assessment while also facilitating efficient generation of F0 animals for subsequent breeding.
Links
Get the data
- GEO FTP directory ftp.ncbi.nlm.nih.gov/geo/series/GSE306nnn/GSE306203 ↗
download · from NCBI GEO
Where it is published
- GEO accession page ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE306203 ↗
landing page · from NCBI GEO
Documentation and papers
- PRJNA1309674 ncbi.nlm.nih.gov/bioproject/PRJNA1309674 ↗
project · from NCBI GEO
- PubMed 42433665 pubmed.ncbi.nlm.nih.gov/42433665 ↗
publication · from NCBI GEO
Topics
- Stated by source
- Expression profiling by high throughput sequencing · Mus musculus
- From keywords
- Life Sciences
- Inferred from text
- RNA sequencing 65% · Single-cell RNA sequencing 65%
Provenance · 1 source records, 9 field assertions
| Source | Key | Last seen | Raw |
|---|---|---|---|
| NCBI GEO | GSE306203 | 14 d ago | JSON v1 |
| Field | Assertion | Extractor | Evidence |
|---|---|---|---|
| access_level | source · NCBI GEO | connector:ncbi_geo@1.0.0 | |
| concepts[field].local:field:life-sciences | mapping · NCBI GEO | connector:ncbi_geo@1.0.0 | |
| concepts[method].geo_series_type:expression-profiling-by-high-throughput-sequencing | source · NCBI GEO | connector:ncbi_geo@1.0.0 | /gdstype |
| concepts[modality].local:modality:rna-seq | enrichment · NCBI GEO | keyword-concept-rules@1.0.0 | title+description (65%) |
| concepts[modality].local:modality:single-cell-rna-seq | enrichment · NCBI GEO | keyword-concept-rules@1.0.0 | title+description (65%) |
| concepts[organism].NCBITaxon:10090 | source · NCBI GEO | connector:ncbi_geo@1.0.0 | /taxon |
| description | source · NCBI GEO | connector:ncbi_geo@1.0.0 | /summary |
| publication_date | source · NCBI GEO | connector:ncbi_geo@1.0.0 | |
| title | source · NCBI GEO | connector:ncbi_geo@1.0.0 | /title |