ArticlePloS one2015
Efficient Generation of Myostatin Knock-Out Sheep Using CRISPR/Cas9 Technology and Microinjection into Zygotes.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 135 papers.
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Who cites it
135 citing papers in PubMed, 307 citations in OpenAlex.
- Non-Coding Negative Regulatory Features in Livestock Genomes: Functional Annotation and Causal Validation.Animals : an open access journal from MDPI · 2026Review
- Precise dual-gene knockout of MSTN and SOCS2 via cytidine base editing enhances muscling and growth in goats.Functional & integrative genomics · 2026Article
- Single-Cell Omics Advances in Understanding Tissue Development and Complex Trait Formation in Sheep and Goats.Animals : an open access journal from MDPI · 2026Review
- Targeted Gene Editing in Honey Bees Using Liposome-Based CRISPR-Cas9.Biochemical genetics · 2026Article
- The origin of the double-muscle mutation in theFrontiers in veterinary science · 2026Article
- Harnessing myostatin pleiotropy for multitrait improvement via precision gene editing.Frontiers in genome editing · 2026Review
- From markers to mechanisms: a comprehensive review of major genes and quantitative trait loci shaping the modern sheep (Frontiers in genetics · 2026Review
- SCNT and genome editing in Buffalo, the black gold of the tropics.The Journal of reproduction and development · 2026Review
- Molecular markers associated with growth, meat, and carcass traits in sheep: a review.Animal biotechnology · 2025Review
- Relationship between the expression levels of myogenic regulatory factor genes and carcass characteristics in Kivircik and Hungarian Merino lambs.Animal biotechnology · 2025Article
- Effects ofAnimals : an open access journal from MDPI · 2025Article
- Cytosine base editor-mediated high-efficiency myostatin editing in Hu sheep.Functional & integrative genomics · 2025Article
- Agricultural biotechnology in China: product development, commercialization, and perspectives.aBIOTECH · 2025Review
- Genetic advancements and future directions in ruminant livestock breeding: from reference genomes to multiomics innovations.Science China. Life sciences · 2025Review
- Generation and propagation of high fecundity gene edited fine wool sheep by CRISPR/Cas9.Scientific reports · 2025Article
- Regulatory Role ofInternational journal of molecular sciences · 2024Article
- Unlocking Genome Editing: Advances and Obstacles in CRISPR/Cas Delivery Technologies.Journal of functional biomaterials · 2024Review
- Electroporation Delivery of Cas9 sgRNA Ribonucleoprotein-Mediated Genome Editing in Sheep IVF Zygotes.International journal of molecular sciences · 2024Article
- Activin Signaling Pathway Specialization During Embryonic and Skeletal Muscle Development in Rainbow Trout (Oncorhynchus mykiss).Marine biotechnology (New York, N.Y.) · 2024Article
- CRISPR-Cas guide RNA indel analysis using CRISPResso2 with Nanopore sequencing data.BMC research notes · 2024Article
75 more citing papers are in PubMed but not listed here.
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Authors and funding
11 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
While CRISPR/Cas9 technology has proven to be a valuable system to generate gene-targeted modified animals in several species, this tool has been scarcely reported in farm animals. Myostatin is encoded by MSTN gene involved in the inhibition of muscle differentiation and growth. We determined the efficiency of the CRISPR/Cas9 system to edit MSTN in sheep and generate knock-out (KO) animals with the aim to promote muscle development and body growth. We generated CRISPR/Cas9 mRNAs specific for ovine MSTN and microinjected them into the cytoplasm of ovine zygotes. When embryo development of CRISPR/Cas9 microinjected zygotes (n = 216) was compared with buffer injected embryos (n = 183) and non microinjected embryos (n = 173), cleavage rate was lower for both microinjected groups (P<0.05) and neither was affected by CRISPR/Cas9 content in the injected medium. Embryo development to blastocyst was not affected by microinjection and was similar among the experimental groups. From 20 embryos analyzed by Sanger sequencing, ten were mutant (heterozygous or mosaic; 50% efficiency). To obtain live MSTN KO lambs, 53 blastocysts produced after zygote CRISPR/Cas9 microinjection were transferred to 29 recipient females resulting in 65.5% (19/29) of pregnant ewes and 41.5% (22/53) of newborns. From 22 born lambs analyzed by T7EI and Sanger sequencing, ten showed indel mutations at MSTN gene. Eight showed mutations in both alleles and five of them were homozygous for indels generating out-of frame mutations that resulted in premature stop codons. Western blot analysis of homozygous KO founders confirmed the absence of myostatin, showing heavier body weight than wild type counterparts. In conclusion, our results demonstrate that CRISPR/Cas9 system was a very efficient tool to generate gene KO sheep. This technology is quick and easy to perform and less expensive than previous techniques, and can be applied to obtain genetically modified animal models of interest for biomedicine and livestock.
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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.