CRISPR/Cas9-editing of PRNP in Alpine goats
Abstract Misfolding of the cellular PrP (PrPc) protein causes prion disease, leading to neurodegenerative disorders in numerous mammalian species, including goats. A lack of PrPc induces complete resistance to prion disease. The aim of this work was to engineer Alpine goats carrying knockout (KO) al...
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BMC
2025-01-01
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| Series: | Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s13567-024-01444-1 |
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| author | Aurélie Allais-Bonnet Christophe Richard Marjolaine André Valérie Gelin Marie-Christine Deloche Aurore Lamadon Gwendoline Morin Béatrice Mandon-Pépin Eugénie Canon Dominique Thépot Johann Laubier Katayoun Moazami-Goudarzi Ludivine Laffont Olivier Dubois Thierry Fassier Patrice Congar Olivier Lasserre Tiphaine Aguirre-Lavin Jean-Luc Vilotte Eric Pailhoux |
| author_facet | Aurélie Allais-Bonnet Christophe Richard Marjolaine André Valérie Gelin Marie-Christine Deloche Aurore Lamadon Gwendoline Morin Béatrice Mandon-Pépin Eugénie Canon Dominique Thépot Johann Laubier Katayoun Moazami-Goudarzi Ludivine Laffont Olivier Dubois Thierry Fassier Patrice Congar Olivier Lasserre Tiphaine Aguirre-Lavin Jean-Luc Vilotte Eric Pailhoux |
| author_sort | Aurélie Allais-Bonnet |
| collection | DOAJ |
| description | Abstract Misfolding of the cellular PrP (PrPc) protein causes prion disease, leading to neurodegenerative disorders in numerous mammalian species, including goats. A lack of PrPc induces complete resistance to prion disease. The aim of this work was to engineer Alpine goats carrying knockout (KO) alleles of PRNP, the PrPc-encoding gene, using CRISPR/Cas9-ribonucleoproteins and single-stranded donor oligonucleotides. The targeted region preceded the PRNP Ter mutation previously described in Norwegian goats. Genome editors were injected under the zona pellucida prior to the electroporation of 565 Alpine goat embryos/oocytes. A total of 122 two-cell-stage embryos were transferred to 46 hormonally synchronized recipient goats. Six of the goats remained pregnant and naturally gave birth to 10 offspring. Among the 10 newborns, eight founder animals carrying PRNP genome-edited alleles were obtained. Eight different mutated alleles were observed, including five inducing KO mutations. Three founders carried only genome-edited alleles and were phenotypically indistinguishable from their wild-type counterparts. Among them, one male carrying a one base pair insertion leading to a KO allele is currently used to rapidly extend a PRNP-KO line of Alpine goats for future characterization. In addition to KO alleles, a PRNP del6 genetic variant has been identified in one-third of founder animals. This new variant will be tested for its potential properties with respect to prion disease. Future studies will also evaluate the effects of genetic background on other characters associated with PRNP KO, as previously described in the Norwegian breed or other species. |
| format | Article |
| id | doaj-art-6107e0fdc8de401193403e30d56b29c4 |
| institution | Kabale University |
| issn | 1297-9716 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Veterinary Research |
| spelling | doaj-art-6107e0fdc8de401193403e30d56b29c42025-01-19T12:35:08ZengBMCVeterinary Research1297-97162025-01-0156111310.1186/s13567-024-01444-1CRISPR/Cas9-editing of PRNP in Alpine goatsAurélie Allais-Bonnet0Christophe Richard1Marjolaine André2Valérie Gelin3Marie-Christine Deloche4Aurore Lamadon5Gwendoline Morin6Béatrice Mandon-Pépin7Eugénie Canon8Dominique Thépot9Johann Laubier10Katayoun Moazami-Goudarzi11Ludivine Laffont12Olivier Dubois13Thierry Fassier14Patrice Congar15Olivier Lasserre16Tiphaine Aguirre-Lavin17Jean-Luc Vilotte18Eric Pailhoux19ElianceUVSQ, INRAE, BREED, Université Paris-SaclayUVSQ, INRAE, BREED, Université Paris-SaclayUVSQ, INRAE, BREED, Université Paris-SaclayElianceUVSQ, INRAE, BREED, Université Paris-SaclayINRAE, SAJUVSQ, INRAE, BREED, Université Paris-SaclayUVSQ, INRAE, BREED, Université Paris-SaclayUVSQ, INRAE, BREED, Université Paris-SaclayINRAE, AgroParisTech, GABI, Université Paris-SaclayINRAE, AgroParisTech, GABI, Université Paris-SaclayUVSQ, INRAE, BREED, Université Paris-SaclayUVSQ, INRAE, BREED, Université Paris-SaclayINRAE, UE P3R Bourges, Domaine de BourgesINRAE, SAJINRAE, PAOINRAE, PAOINRAE, AgroParisTech, GABI, Université Paris-SaclayUVSQ, INRAE, BREED, Université Paris-SaclayAbstract Misfolding of the cellular PrP (PrPc) protein causes prion disease, leading to neurodegenerative disorders in numerous mammalian species, including goats. A lack of PrPc induces complete resistance to prion disease. The aim of this work was to engineer Alpine goats carrying knockout (KO) alleles of PRNP, the PrPc-encoding gene, using CRISPR/Cas9-ribonucleoproteins and single-stranded donor oligonucleotides. The targeted region preceded the PRNP Ter mutation previously described in Norwegian goats. Genome editors were injected under the zona pellucida prior to the electroporation of 565 Alpine goat embryos/oocytes. A total of 122 two-cell-stage embryos were transferred to 46 hormonally synchronized recipient goats. Six of the goats remained pregnant and naturally gave birth to 10 offspring. Among the 10 newborns, eight founder animals carrying PRNP genome-edited alleles were obtained. Eight different mutated alleles were observed, including five inducing KO mutations. Three founders carried only genome-edited alleles and were phenotypically indistinguishable from their wild-type counterparts. Among them, one male carrying a one base pair insertion leading to a KO allele is currently used to rapidly extend a PRNP-KO line of Alpine goats for future characterization. In addition to KO alleles, a PRNP del6 genetic variant has been identified in one-third of founder animals. This new variant will be tested for its potential properties with respect to prion disease. Future studies will also evaluate the effects of genetic background on other characters associated with PRNP KO, as previously described in the Norwegian breed or other species.https://doi.org/10.1186/s13567-024-01444-1PRNP genePRION proteingoatanimal healthembryo editingknockout |
| spellingShingle | Aurélie Allais-Bonnet Christophe Richard Marjolaine André Valérie Gelin Marie-Christine Deloche Aurore Lamadon Gwendoline Morin Béatrice Mandon-Pépin Eugénie Canon Dominique Thépot Johann Laubier Katayoun Moazami-Goudarzi Ludivine Laffont Olivier Dubois Thierry Fassier Patrice Congar Olivier Lasserre Tiphaine Aguirre-Lavin Jean-Luc Vilotte Eric Pailhoux CRISPR/Cas9-editing of PRNP in Alpine goats Veterinary Research PRNP gene PRION protein goat animal health embryo editing knockout |
| title | CRISPR/Cas9-editing of PRNP in Alpine goats |
| title_full | CRISPR/Cas9-editing of PRNP in Alpine goats |
| title_fullStr | CRISPR/Cas9-editing of PRNP in Alpine goats |
| title_full_unstemmed | CRISPR/Cas9-editing of PRNP in Alpine goats |
| title_short | CRISPR/Cas9-editing of PRNP in Alpine goats |
| title_sort | crispr cas9 editing of prnp in alpine goats |
| topic | PRNP gene PRION protein goat animal health embryo editing knockout |
| url | https://doi.org/10.1186/s13567-024-01444-1 |
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