Endangered Przewalski’s Horse, <i>Equus przewalskii</i>, Cloned from Historically Cryopreserved Cells

Cloning from historically cryopreserved cells offers a potential means to restore lost genetic variation or increase the representation of particular lineages within bottlenecked species, provided such biobanked materials are archived for such genetic rescue applications. One species for which cloni...

Full description

Saved in:
Bibliographic Details
Main Authors: Ben J. Novak, Oliver A. Ryder, Marlys L. Houck, Kelcey Walker, Lexie Russell, Blake Russell, Shawn Walker, Sanaz Sadeghieh Arenivas, Lauren Aston, Gregg Veneklasen, Jamie A. Ivy, Klaus-Peter Koepfli, Anna Rusnak, Jaroslav Simek, Anna Zhuk, Andrea S. Putnam, Ryan Phelan
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Animals
Subjects:
Online Access:https://www.mdpi.com/2076-2615/15/5/613
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850030907481653248
author Ben J. Novak
Oliver A. Ryder
Marlys L. Houck
Kelcey Walker
Lexie Russell
Blake Russell
Shawn Walker
Sanaz Sadeghieh Arenivas
Lauren Aston
Gregg Veneklasen
Jamie A. Ivy
Klaus-Peter Koepfli
Anna Rusnak
Jaroslav Simek
Anna Zhuk
Andrea S. Putnam
Ryan Phelan
author_facet Ben J. Novak
Oliver A. Ryder
Marlys L. Houck
Kelcey Walker
Lexie Russell
Blake Russell
Shawn Walker
Sanaz Sadeghieh Arenivas
Lauren Aston
Gregg Veneklasen
Jamie A. Ivy
Klaus-Peter Koepfli
Anna Rusnak
Jaroslav Simek
Anna Zhuk
Andrea S. Putnam
Ryan Phelan
author_sort Ben J. Novak
collection DOAJ
description Cloning from historically cryopreserved cells offers a potential means to restore lost genetic variation or increase the representation of particular lineages within bottlenecked species, provided such biobanked materials are archived for such genetic rescue applications. One species for which cloning can provide genetic management benefits is Przewalski’s horse, <i>Equus przewalskii</i>. All ~1800 living Przewalski’s horses, distributed across ex situ breeding facilities and in situ reintroduction sites, are descended from one or more of the five founder lineages established by the 12 horses captured from the wild between 1898 and 1947. Since the 1970s, the San Diego Zoo Wildlife Alliance Biodiversity Bank’s Frozen Zoo<sup>®</sup> (Escondido, CA, USA) has biobanked cells or tissues of 575 individuals spanning many generations. A pedigree analysis of a subset of deceased individuals represented in the San Diego Zoo Wildlife Alliance Biodiversity Bank’s Frozen Zoo<sup>®</sup> revealed an underrepresented male that lived from 1975 to 1998, Studbook Number 615 (also known as Kuporovitch), who would be of high value for breeding if cloned. Here, we report that two healthy clones were produced from this cell line using cross-species somatic cell nuclear transfer from 2020 to 2023. Their identification as clones was verified by a standard horse-pedigree genotyping panel, and, for one clone, a whole genome sequencing comparison to the original donor was performed. This is the first time that multiple healthy clones surviving the perinatal period have been produced for an endangered species.
format Article
id doaj-art-3d1626bde4424bd68aa6f8a4de8efe9c
institution DOAJ
issn 2076-2615
language English
publishDate 2025-02-01
publisher MDPI AG
record_format Article
series Animals
spelling doaj-art-3d1626bde4424bd68aa6f8a4de8efe9c2025-08-20T02:59:07ZengMDPI AGAnimals2076-26152025-02-0115561310.3390/ani15050613Endangered Przewalski’s Horse, <i>Equus przewalskii</i>, Cloned from Historically Cryopreserved CellsBen J. Novak0Oliver A. Ryder1Marlys L. Houck2Kelcey Walker3Lexie Russell4Blake Russell5Shawn Walker6Sanaz Sadeghieh Arenivas7Lauren Aston8Gregg Veneklasen9Jamie A. Ivy10Klaus-Peter Koepfli11Anna Rusnak12Jaroslav Simek13Anna Zhuk14Andrea S. Putnam15Ryan Phelan16Revive & Restore, 1505 Bridgeway #203, Sausalito, CA 94965, USABeckman Center for Conservation Research, San Diego Zoo Wildlife Alliance, Escondido, CA 92027, USABeckman Center for Conservation Research, San Diego Zoo Wildlife Alliance, Escondido, CA 92027, USAViaGen Pets & Equine, 715 Discovery Blvd #410, Cedar Park, TX 78613, USAViaGen Pets & Equine, 715 Discovery Blvd #410, Cedar Park, TX 78613, USAViaGen Pets & Equine, 715 Discovery Blvd #410, Cedar Park, TX 78613, USAViaGen Pets & Equine, 715 Discovery Blvd #410, Cedar Park, TX 78613, USAViaGen Pets & Equine, 715 Discovery Blvd #410, Cedar Park, TX 78613, USAViaGen Pets & Equine, 715 Discovery Blvd #410, Cedar Park, TX 78613, USATimber Creek Veterinary Hospital, 19302 Farm to Market Rd 1541, Canyon, TX 79015, USAIndependent Researcher, Erie, CO 80516, USASmithsonian-Mason School of Conservation, George Mason University, 1500 Remount Road, Front Royal, VA 22630, USAInstitute of Applied Computer Science, ITMO University, 197101 St. Petersburg, RussiaPrague Zoo, 171 00 Prague, Czech RepublicInstitute of Applied Computer Science, ITMO University, 197101 St. Petersburg, RussiaDallas Zoo, 650 S R.L. Thornton Fwy, Dallas, TX 75203, USARevive & Restore, 1505 Bridgeway #203, Sausalito, CA 94965, USACloning from historically cryopreserved cells offers a potential means to restore lost genetic variation or increase the representation of particular lineages within bottlenecked species, provided such biobanked materials are archived for such genetic rescue applications. One species for which cloning can provide genetic management benefits is Przewalski’s horse, <i>Equus przewalskii</i>. All ~1800 living Przewalski’s horses, distributed across ex situ breeding facilities and in situ reintroduction sites, are descended from one or more of the five founder lineages established by the 12 horses captured from the wild between 1898 and 1947. Since the 1970s, the San Diego Zoo Wildlife Alliance Biodiversity Bank’s Frozen Zoo<sup>®</sup> (Escondido, CA, USA) has biobanked cells or tissues of 575 individuals spanning many generations. A pedigree analysis of a subset of deceased individuals represented in the San Diego Zoo Wildlife Alliance Biodiversity Bank’s Frozen Zoo<sup>®</sup> revealed an underrepresented male that lived from 1975 to 1998, Studbook Number 615 (also known as Kuporovitch), who would be of high value for breeding if cloned. Here, we report that two healthy clones were produced from this cell line using cross-species somatic cell nuclear transfer from 2020 to 2023. Their identification as clones was verified by a standard horse-pedigree genotyping panel, and, for one clone, a whole genome sequencing comparison to the original donor was performed. This is the first time that multiple healthy clones surviving the perinatal period have been produced for an endangered species.https://www.mdpi.com/2076-2615/15/5/613somatic cell nuclear transfercloninggenetic rescuePrzewalski’s horsebiobanking
spellingShingle Ben J. Novak
Oliver A. Ryder
Marlys L. Houck
Kelcey Walker
Lexie Russell
Blake Russell
Shawn Walker
Sanaz Sadeghieh Arenivas
Lauren Aston
Gregg Veneklasen
Jamie A. Ivy
Klaus-Peter Koepfli
Anna Rusnak
Jaroslav Simek
Anna Zhuk
Andrea S. Putnam
Ryan Phelan
Endangered Przewalski’s Horse, <i>Equus przewalskii</i>, Cloned from Historically Cryopreserved Cells
Animals
somatic cell nuclear transfer
cloning
genetic rescue
Przewalski’s horse
biobanking
title Endangered Przewalski’s Horse, <i>Equus przewalskii</i>, Cloned from Historically Cryopreserved Cells
title_full Endangered Przewalski’s Horse, <i>Equus przewalskii</i>, Cloned from Historically Cryopreserved Cells
title_fullStr Endangered Przewalski’s Horse, <i>Equus przewalskii</i>, Cloned from Historically Cryopreserved Cells
title_full_unstemmed Endangered Przewalski’s Horse, <i>Equus przewalskii</i>, Cloned from Historically Cryopreserved Cells
title_short Endangered Przewalski’s Horse, <i>Equus przewalskii</i>, Cloned from Historically Cryopreserved Cells
title_sort endangered przewalski s horse i equus przewalskii i cloned from historically cryopreserved cells
topic somatic cell nuclear transfer
cloning
genetic rescue
Przewalski’s horse
biobanking
url https://www.mdpi.com/2076-2615/15/5/613
work_keys_str_mv AT benjnovak endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT oliveraryder endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT marlyslhouck endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT kelceywalker endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT lexierussell endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT blakerussell endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT shawnwalker endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT sanazsadeghieharenivas endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT laurenaston endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT greggveneklasen endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT jamieaivy endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT klauspeterkoepfli endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT annarusnak endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT jaroslavsimek endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT annazhuk endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT andreasputnam endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells
AT ryanphelan endangeredprzewalskishorseiequusprzewalskiiiclonedfromhistoricallycryopreservedcells