ArticleThe Journal of reproduction and development2017
Production of α1,3-galactosyltransferase and cytidine monophosphate-N-acetylneuraminic acid hydroxylase gene double-deficient pigs by CRISPR/Cas9 and handmade cloning.
Article in The Journal of reproduction and development, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- CRISPR-Cas9 in the Tailoring of Genetically Engineered Animals.Current issues in molecular biology · 2025Review
- Claudin-2 enhances human antibody-mediated complement-dependent cytotoxicity of porcine endothelial cells by modulating antibody binding and complement activation.Frontiers in immunology · 2025Article
- Review
- Advances in pig models of human diseases.Animal models and experimental medicine · 2022Review
- Genetically engineered pigs for xenotransplantation: Hopes and challenges.Frontiers in cell and developmental biology · 2022Review
- Human IL-17 and TNF-α Additively or Synergistically Regulate the Expression of Proinflammatory Genes, Coagulation-Related Genes, and Tight Junction Genes in Porcine Aortic Endothelial Cells.Frontiers in immunology · 2022Article
- Advances in CRISPR-Based Functional Genomics and Nucleic Acid Detection in Pigs.Frontiers in genetics · 2022Article
- Application of CRISPR/Cas9 System in Establishing Large Animal Models.Frontiers in cell and developmental biology · 2022Review
- Current status of the application of gene editing in pigs.The Journal of reproduction and development · 2021Review
- CRISPR/Cas Technology in Pig-to-Human Xenotransplantation Research.International journal of molecular sciences · 2021Review
- Efficient generation of GGTA1-deficient pigs by electroporation of the CRISPR/Cas9 system into in vitro-fertilized zygotes.BMC biotechnology · 2020Article
- The Possible Role of Anti-Neu5Gc as an Obstacle in Xenotransplantation.Frontiers in immunology · 2020Review
- Improvements in Gene Editing Technology Boost Its Applications in Livestock.Frontiers in genetics · 2020Review
- TNF-α promotes human antibody-mediated complement-dependent cytotoxicity of porcine endothelial cells through downregulating P38-mediated Occludin expression.Cell communication and signaling : CCS · 2019Article
- Reducing porcine corneal graft rejection, with an emphasis on porcine endogenous retrovirus transmission safety: a review.International journal of ophthalmology · 2019Review
- Article
- Evolution of sialic acids: Implications in xenotransplant biology.Xenotransplantation · 2018Review
- Will Genetic Engineering Carry Xenotransplantation of Pig Islets to the Clinic?Current diabetes reports · 2018Review
- [Research progress of producing genetically modified pigs by CRISPR/Cas9 in the medical field].Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi · 2018Review
- Genome Editing of Pigs for Agriculture and Biomedicine.Frontiers in genetics · 2018Review
Corrections and comments
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Authors and funding
12 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Gene-knockout pigs hold great promise as a solution to the shortage of organs from donor animals for xenotransplantation. Several groups have generated gene-knockout pigs via clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) and somatic cell nuclear transfer (SCNT). Herein, we adopted a simple and micromanipulator-free method, handmade cloning (HMC) instead of SCNT, to generate double gene-knockout pigs. First, we applied the CRISPR/Cas9 system to target α1,3-galactosyltransferase (GGTA1) and cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) genes simultaneously in porcine fetal fibroblast cells (PFFs), which were derived from wild-type Chinese domestic miniature Wuzhishan pigs. Cell colonies were obtained by screening and were identified by Surveyor assay and sequencing. Next, we chose the GGTA1/CMAH double-knockout (DKO) cells for HMC to produce piglets. As a result, we obtained 11 live bi-allelic GGTA1/CMAH DKO piglets with the identical phenotype. Compared to cells from GGTA1-knockout pigs, human antibody binding and antibody-mediated complement-dependent cytotoxicity were significantly reduced in cells from GGTA1/CMAH DKO pigs, which demonstrated that our pigs would exhibit reduced humoral rejection in xenotransplantation. These data suggested that the combination of CRISPR/Cas9 and HMC technology provided an efficient and new strategy for producing pigs with multiple genetic modifications.
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