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פותח על ידי קלירמאש פתרונות בע"מ -
Gene augmentation therapy for a missense substitution in the cGMP-binding domain of ovine CNGA3 gene restores vision in day-blind sheep
Year:
2017
Authors :
ניצן, טלי
;
.
Volume :
58
Co-Authors:
Gootwine, E., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Abu-Siam, M., Abu Siam Veterinary Clinic, Rahat, Israel
Obolensky, A., Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
Rosov, A., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Honig, H., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Nitzan, T., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Shirak, A., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Ezra-Elia, R., Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel
Yamin, E., Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
Banin, E., Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
Averbukh, E., Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
Hauswirth, W.W., Department of Ophthalmology, University of Florida, Gainesville, FL, United States
Ofri, R., Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel
Seroussi, E., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Facilitators :
From page:
1577
To page:
1584
(
Total pages:
8
)
Abstract:
Purpose. Applying CNGA3 gene augmentation therapy to cure a novel causative mutation underlying achromatopsia (ACHM) in sheep. Methods. Impaired vision that spontaneously appeared in newborn lambs was characterized by behavioral, electroretinographic (ERG), and histologic techniques. Deep-sequencing reads of an affected lamb and an unaffected lamb were compared within conserved genomic regions orthologous to human genes involved in similar visual impairment. Observed nonsynonymous amino acid substitutions were classified by their deleteriousness score. The putative causative mutation was assessed by producing compound CNGA3 heterozygotes and applying gene augmentation therapy using the orthologous human cDNA. Results. Behavioral assessment revealed day blindness, and subsequent ERG examination showed attenuated photopic responses. Histologic and immunohistochemical examination of affected sheep eyes did not reveal degeneration, and cone photoreceptors expressing CNGA3 were present. Bioinformatics and sequencing analyses suggested a c.1618G>A, p.Gly540Ser substitution in the GMP-binding domain of CNGA3 as the causative mutation. This was confirmed by genetic concordance test and by genetic complementation experiment: All five compound CNGA3 heterozygotes, carrying both p.Arg236* and p.Gly540Ser mutations in CNGA3, were day-blind. Furthermore, subretinal delivery of the intact human CNGA3 gene using an adeno-associated viral vector (AAV) restored photopic vision in two affected p.Gly540Ser homozygous rams. Conclusions. The c.1618G>A, p.Gly540Ser substitution in CNGA3 was identified as the causative mutation for a novel form of ACHM in Awassi sheep. Gene augmentation therapy restored vision in the affected sheep. This novel mutation provides a large-animal model that is valid for most human CNGA3 ACHM patients; the majority of them carry missense rather than premature-termination mutations. © 2017 The Authors.
Note:
Related Files :
Animals
Awassi
Female
gene therapy
Genetics
Male
mutation
sheep
עוד תגיות
תוכן קשור
More details
DOI :
10.1167/iovs.16-20986
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
21805
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:46
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Scientific Publication
Gene augmentation therapy for a missense substitution in the cGMP-binding domain of ovine CNGA3 gene restores vision in day-blind sheep
58
Gootwine, E., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Abu-Siam, M., Abu Siam Veterinary Clinic, Rahat, Israel
Obolensky, A., Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
Rosov, A., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Honig, H., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Nitzan, T., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Shirak, A., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Ezra-Elia, R., Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel
Yamin, E., Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
Banin, E., Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
Averbukh, E., Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
Hauswirth, W.W., Department of Ophthalmology, University of Florida, Gainesville, FL, United States
Ofri, R., Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel
Seroussi, E., Agricultural Research Organization, The Volcani Center, Rishon LeZion, Israel
Gene augmentation therapy for a missense substitution in the cGMP-binding domain of ovine CNGA3 gene restores vision in day-blind sheep
Purpose. Applying CNGA3 gene augmentation therapy to cure a novel causative mutation underlying achromatopsia (ACHM) in sheep. Methods. Impaired vision that spontaneously appeared in newborn lambs was characterized by behavioral, electroretinographic (ERG), and histologic techniques. Deep-sequencing reads of an affected lamb and an unaffected lamb were compared within conserved genomic regions orthologous to human genes involved in similar visual impairment. Observed nonsynonymous amino acid substitutions were classified by their deleteriousness score. The putative causative mutation was assessed by producing compound CNGA3 heterozygotes and applying gene augmentation therapy using the orthologous human cDNA. Results. Behavioral assessment revealed day blindness, and subsequent ERG examination showed attenuated photopic responses. Histologic and immunohistochemical examination of affected sheep eyes did not reveal degeneration, and cone photoreceptors expressing CNGA3 were present. Bioinformatics and sequencing analyses suggested a c.1618G>A, p.Gly540Ser substitution in the GMP-binding domain of CNGA3 as the causative mutation. This was confirmed by genetic concordance test and by genetic complementation experiment: All five compound CNGA3 heterozygotes, carrying both p.Arg236* and p.Gly540Ser mutations in CNGA3, were day-blind. Furthermore, subretinal delivery of the intact human CNGA3 gene using an adeno-associated viral vector (AAV) restored photopic vision in two affected p.Gly540Ser homozygous rams. Conclusions. The c.1618G>A, p.Gly540Ser substitution in CNGA3 was identified as the causative mutation for a novel form of ACHM in Awassi sheep. Gene augmentation therapy restored vision in the affected sheep. This novel mutation provides a large-animal model that is valid for most human CNGA3 ACHM patients; the majority of them carry missense rather than premature-termination mutations. © 2017 The Authors.
Scientific Publication
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