ArticleScientific reports2016
Isozygous and selectable marker-free MSTN knockout cloned pigs generated by the combined use of CRISPR/Cas9 and Cre/LoxP.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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36 citing papers in PubMed, 92 citations in OpenAlex.
- Effects ofAnimals : an open access journal from MDPI · 2025Article
- A bibliometric mapping of advancements and trends in genome editing in pigs.Tropical animal health and production · 2025Review
- Molecular breeding of pigs in the genome editing era.Genetics, selection, evolution : GSE · 2025Review
- CRISPR/Cas9-TargetedJournal of developmental biology · 2025Article
- Combined metabolome and transcriptome analysis revealed that MSTN regulated the process of bovine fatty acid metabolism in gut.Frontiers in veterinary science · 2025Article
- CRISPR Technology Acts as a Dual-Purpose Tool in Pig Breeding: Enhancing Both Agricultural Productivity and Biomedical Applications.Biomolecules · 2024Review
- Precise Insertion of AttB Sequences in Goat Genome Using Enhanced Prime Editor.International journal of molecular sciences · 2024Article
- Myostatin gene role in regulating traits of poultry species for potential industrial applications.Journal of animal science and biotechnology · 2024Review
- Current status and future of gene engineering in livestock.BMB reports · 2024Review
- Efficient and Specific Generation ofGenes · 2023Article
- Myostatin increases the expression of matrix metalloproteinase genes to promote preadipocytes differentiation in pigs.Adipocyte · 2022Article
- Potentials, prospects and applications of genome editing technologies in livestock production.Saudi journal of biological sciences · 2022Review
- Increased sizes and improved qualities of tibia bones by myostatin mutation in Japanese quail.Frontiers in physiology · 2022Article
- Generation of Heritable Prominent Double Muscle Buttock Rabbits via Novel Site Editing of Myostatin Gene Using CRISPR/Cas9 System.Frontiers in veterinary science · 2022Article
- Myostatin Mutation in Japanese Quail Increased Egg Size but Reduced Eggshell Thickness and Strength.Animals : an open access journal from MDPI · 2021Article
- Generation of mutant pigs by lipofection-mediated genome editing in embryos.Scientific reports · 2021Article
- Porcine germline genome engineering.Proceedings of the National Academy of Sciences of the United States of America · 2021Review
- Insulin-Degrading Enzyme Regulates the Proliferation and Apoptosis of Porcine Skeletal Muscle Stem CellsFrontiers in cell and developmental biology · 2021Article
- Adaptation of Gut Microbiome to Transgenic Pigs Secreting β-Glucanase, Xylanase, and Phytase.Frontiers in genetics · 2021Article
- Efficient CRISPR/Cas9-mediated gene editing in Guangdong small-ear spotted pig cells using an optimized electrotransfection method.Biotechnology letters · 2020Article
Corrections and comments
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Authors and funding
13 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Predictable, clean genetic modification (GM) in livestock is important for reliable phenotyping and biosafety. Here we reported the generation of isozygous, functional myostatin (MSTN) knockout cloned pigs free of selectable marker gene (SMG) by CRISPR/Cas9 and Cre/LoxP. CRISPR/Cas9-mediated homologous recombination (HR) was exploited to knock out (KO) one allele of MSTN in pig primary cells. Cre recombinase was then used to excise the SMG with an efficiency of 82.7%. The SMG-free non-EGFP cells were isolated by flow cytometery and immediately used as donor nuclei for nuclear transfer. A total of 685 reconstructed embryos were transferred into three surrogates with one delivering two male live piglets. Molecular testing verified the mono-allelic MSTN KO and SMG deletion in these cloned pigs. Western blots showed approximately 50% decrease in MSTN and concurrent increased expression of myogenic genes in muscle. Histological examination revealed the enhanced myofiber quantity but myofiber size remained unaltered. Ultrasonic detection showed the increased longissimus muscle size and decreased backfat thickness. Precision editing of pig MSTN gene has generated isozygous, SMG-free MSTN KO cloned founders, which guaranteed a reliable route for elite livestock production and a strategy to minimize potential biological risks.
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