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Generation of cattle knockout for galactose‐α1,3‐galactose and N‐glycolylneuraminic acid antigens
- Source :
- Xenotransplantation, Xenotransplantation, Wiley, 2019, pp.e12524. ⟨10.1111/xen.12524⟩, Xenotransplantation, 2019, 26 (5), pp.e12524. ⟨10.1111/xen.12524⟩, Xenotransplantation, Wiley, 2019, 26 (5), pp.e12524. ⟨10.1111/xen.12524⟩, Dipòsit Digital de la UB, Universidad de Barcelona, Dipòsit Digital de Documents de la UAB, Universitat Autònoma de Barcelona
- Publication Year :
- 2019
- Publisher :
- HAL CCSD, 2019.
-
Abstract
- International audience; Two well-characterized carbohydrate epitopes are absent in humans but present in other mammals. These are galactose-α1,3-galactose (αGal) and N-glycolylneuraminic acid (Neu5Gc) which are introduced by the activities of two enzymes including α(1,3) galactosyltransferase (encoded by the GGTA1 gene) and CMP-Neu5Gc hydroxylase (encoded by the CMAH gene) that are inactive in humans but present in cattle. Hence, bovine-derived products are antigenic in humans who receive bioprosthetic heart valves (BHVs) or those that suffer from red meat syndrome. Using programmable nucleases, we disrupted (knockout, KO) GGTA1 and CMAH genes encoding for the enzymes that catalyse the synthesis of αGal and Neu5Gc, respectively, in both male and female bovine fibroblasts. The KO in clonally selected fibroblasts was detected by polymerase chain reaction (PCR) and confirmed by Sanger sequencing. Selected fibroblasts colonies were used for somatic cell nuclear transfer (SCNT) to produce cloned embryos that were implanted in surrogate recipient heifers. Fifty-three embryos were implanted in 33 recipients heifers; 3 pregnancies were carried to term and delivered 3 live calves. Primary cell cultures were established from the 3 calves and following molecular analyses confirmed the genetic deletions. FACS analysis showed the double-KO phenotype for both antigens confirming the mutated genotypes. Availability of such cattle double-KO model lacking both αGal and Neu5Gc offers a unique opportunity to study the functionality of BHV manufactured with tissues of potentially lower immunogenicity, as well as a possible new clinical approaches to help patients with red meat allergy syndrome due to the presence of these xenoantigens in the diet.
- Subjects :
- Male
0301 basic medicine
knockout
Neu5Gc
030230 surgery
Epitope
Mixed Function Oxygenases
law.invention
Animals, Genetically Modified
Gene Knockout Techniques
chemistry.chemical_compound
0302 clinical medicine
Carn de boví
N-Glycolylneuraminic acid
law
xenotransplantation
Polymerase chain reaction
2. Zero hunger
Sanger sequencing
Heart valves
[SDV.MHEP] Life Sciences [q-bio]/Human health and pathology
Immunogenicity
Galactosyltransferases
Vàlvules cardíaques
Heart Valve Prosthesis
symbols
bioprosthetic Heart Valve (BHV)
Somatic cell nuclear transfer
Xenotransplantation
Female
Original Article
Beef
cattle
CMAH
GGTA1
αGal
Food Hypersensitivity
Knockout
Transplantation, Heterologous
Immunology
Biology
03 medical and health sciences
symbols.namesake
Antigens, Heterophile
Cytidine Monophosphate
Animals
Humans
Gene
Bioprosthesis
Galactosyltransferase
Transplantation
Galactose
Original Articles
Fibroblasts
Bioprosthetic Heart Valve (BHV)
Molecular biology
030104 developmental biology
chemistry
Cattle
Neuraminic Acids
[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Subjects
Details
- Language :
- English
- ISSN :
- 0908665X and 13993089
- Database :
- OpenAIRE
- Journal :
- Xenotransplantation, Xenotransplantation, Wiley, 2019, pp.e12524. ⟨10.1111/xen.12524⟩, Xenotransplantation, 2019, 26 (5), pp.e12524. ⟨10.1111/xen.12524⟩, Xenotransplantation, Wiley, 2019, 26 (5), pp.e12524. ⟨10.1111/xen.12524⟩, Dipòsit Digital de la UB, Universidad de Barcelona, Dipòsit Digital de Documents de la UAB, Universitat Autònoma de Barcelona
- Accession number :
- edsair.doi.dedup.....b4451759e2e6f7c56871bf4ebfd8dd77