ArticleBMC biology2022
Highly reliable creation of floxed alleles by electroporating single-cell embryos.
Article in BMC biology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 15 citations in OpenAlex.
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- BPTF is essential for vaccine-induced germinal center B cell responses.Journal of immunology (Baltimore, Md. : 1950) · 2026Article
- Efficient multi-kilobase knock-ins in mice and cell lines using CRISPR/Cas9 and rAAV donors with unbiased whole-genome characterization by LOCK-seq.Nucleic acids research · 2026Article
- The long noncoding RNAbioRxiv : the preprint server for biology · 2025Article
- Macrophage-T cell interactions promote SLAMF1 expression for enhanced TB defense.Nature communications · 2025Article
- Generation of Plexin-B1 Conditional Knockout Mouse With CRISPR/Cas9 Technology.Genesis (New York, N.Y. : 2000) · 2025Article
- Efficient genome editing of two-cell mouse embryos via modified CRISPR/Cas electroporation.Scientific reports · 2024Article
- West Nile Virus-Induced Expression of Senescent GeneBiomolecules · 2024Article
- Lrp1 is essential for lethal Rift Valley fever hepatic disease in mice.Science advances · 2023Article
- Differential regulation of cardiac sodium channels by intracellular fibroblast growth factors.The Journal of general physiology · 2023Article
- Floxing by Electroporating Single-Cell Embryos with Two CRISPR RNPs and Two ssODNs.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Loss of Stathmin-2, a hallmark of TDP-43-associated ALS, causes motor neuropathy.Cell reports · 2022Article
- Overcoming the bottleneck to widespread testing: a rapid review of nucleic acid testing approaches for COVID-19 detection.RNA (New York, N.Y.) · 2020Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
backgroundFloxed (flanked by loxP) alleles are a crucial portion of conditional knockout mouse models. However, an efficient and reliable strategy to flox genomic regions of any desired size is still lacking.
resultsHere, we demonstrate that the method combining electroporation of fertilized eggs with gRNA/Cas9 complexes and single-stranded oligodeoxynucleotides (ssODNs), assessing phasing of loxP insertions in founders using an in vitro Cre assay and an optional, highly specific and efficient second-round targeting ensures the generation of floxed F1 animals in roughly five months for a wide range of sequence lengths (448 bp to 160 kb reported here).
conclusionsFloxed alleles can be reliably obtained in a predictable timeline using the improved method of electroporation of two gRNA/Cas9 ribonucleoprotein particles (RNPs) and two ssODNs.
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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.