ArticleJournal of animal science and biotechnology2023
Generation of double knockout cattle via CRISPR-Cas9 ribonucleoprotein (RNP) electroporation.
Article in Journal of animal science and biotechnology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 32 citations in OpenAlex.
- Electroporation-mediated site-specific knock-in of a human coding sequence into the bovine ALB locus in embryos.BMC biotechnology · 2026Article
- Production of gene-edited cloned cattle embryos using the CRISPR/EOCas12i system.Frontiers in genome editing · 2026Article
- Empirical optimization of dual-sgRNA design for in vivo CRISPR/Cas9-mediated exon deletion in mice.Computational and structural biotechnology journal · 2026Article
- Genome edited sheep generated by zygote electroporation of prime editor ribonucleoproteins.Frontiers in genome editing · 2026Article
- Harnessing myostatin pleiotropy for multitrait improvement via precision gene editing.Frontiers in genome editing · 2026Review
- Validation of caprine H11 and the Rosa26 platform for transgene integration via CRISPR-based system: investigations on stable transgene expression and genetic biosafety.Functional & integrative genomics · 2025Article
- CRISPR-Cas9 in the Tailoring of Genetically Engineered Animals.Current issues in molecular biology · 2025Review
- MSTN knockout enhances the production of MYOD1-mediated steak-type cultivated meat.Journal of animal science and biotechnology · 2025Article
- Comparative Analysis of Methods for Assessing On-Target Gene Editing Efficiencies.Methods and protocols · 2025Article
- CRISPR/Cas9-editing of PRNP in Alpine goats.Veterinary research · 2025Article
- Long-term (10-year) monitoring of transposon-mediated transgenic cattle.Transgenic research · 2024Article
- An overview of recent technological developments in bovine genomics.Veterinary and animal science · 2024Review
- Current status and future of gene engineering in livestock.BMB reports · 2024Review
Corrections and comments
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Authors and funding
13 authors at 3 institutions in 2 countries.
Funding
Abstract
backgroundGenome editing has been considered as powerful tool in agricultural fields. However, genome editing progress in cattle has not been fast as in other mammal species, for some disadvantages including long gestational periods, single pregnancy, and high raising cost. Furthermore, technically demanding methods such as microinjection and somatic cell nuclear transfer (SCNT) are needed for gene editing in cattle. In this point of view, electroporation in embryos has been risen as an alternative.
resultsFirst, editing efficiency of our electroporation methods were tested for embryos. Presence of mutation on embryo was confirmed by T7E1 assay. With first combination, mutation rates for MSTN and PRNP were 57.6% ± 13.7% and 54.6% ± 13.5%, respectively. In case of MSTN/BLG, mutation rates were 83.9% ± 23.6% for MSTN, 84.5% ± 18.0% for BLG. Afterwards, the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep sequencing. Thirteen recipients were transferred for MSTN/PRNP, 4 calves were delivered, and one calf underwent an induction for double KO. Ten surrogates were given double-KO embryos for MSTN/BLG, and four of the six calves that were born had mutations in both genes.
conclusionsThese data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively applied. Finally, MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.
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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.