Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity

Qiantao Zheng, Jun Lin, Jiaojiao Huang, Hongyong Zhang, Rui Zhang, Xueying Zhang, Chunwei Cao, Catherine Hambly, Guosong Qin, Jing Yao, Ruigao Song, Qitao Jia, Xiao Wang, Yongshun Li, Nan Zhang, Zhengyu Piao, Rongcai Ye, John R Speakman, Hongmei Wang, Qi ZhouYanfang Wang, Wanzhu Jin, Jianguo Zhao

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Abstract

Uncoupling protein 1 (UCP1) is localized on the inner mitochondrial membrane and generates heat by uncoupling ATP synthesis from proton transit across the inner membrane. UCP1 is a key element of nonshivering thermogenesis and is most likely important in the regulation of body adiposity. Pigs (Artiodactyl family Suidae) lack a functional UCP1 gene, resulting in poor thermoregulation and susceptibility to cold, which is an economic and pig welfare issue owing to neonatal mortality. Pigs also have a tendency toward fat accumulation, which may be linked to their lack of UCP1, and thus influences the efficiency of pig production. Here, we report application of a CRISPR/Cas9-mediated, homologous recombination (HR)-independent approach to efficiently insert mouse adiponectin-UCP1 into the porcine endogenous UCP1 locus. The resultant UCP1 knock-in (KI) pigs showed an improved ability to maintain body temperature during acute cold exposure, but they did not have alterations in physical activity levels or total daily energy expenditure (DEE). Furthermore, ectopic UCP1 expression in white adipose tissue (WAT) dramatically decreased fat deposition by 4.89% (P < 0.01), consequently increasing carcass lean percentage (CLP; P < 0.05). Mechanism studies indicated that the loss of fat upon UCP1 activation in WAT was linked to elevated lipolysis. UCP1 KI pigs are a potentially valuable resource for agricultural production through their combination of cold adaptation, which improves pig welfare and reduces economic losses, with reduced fat deposition and increased lean meat production.

Original languageEnglish
Pages (from-to)9474-9482
Number of pages9
JournalPNAS
Volume114
Issue number45
Early online date23 Oct 2017
DOIs
Publication statusPublished - 2017

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Clustered Regularly Interspaced Short Palindromic Repeats
White Adipose Tissue
Swine
Fats
Economics
Uncoupling Protein 1
Thermogenesis
Body Temperature Regulation
Lipolysis
Homologous Recombination
Adiposity
Mitochondrial Membranes
Infant Mortality
Body Temperature
Meat
Energy Metabolism
Protons

Keywords

  • Journal Article
  • UCP1
  • thermoregulation
  • fat deposition
  • pig
  • CRISPR/Cas9

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Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity. / Zheng, Qiantao; Lin, Jun; Huang, Jiaojiao; Zhang, Hongyong; Zhang, Rui; Zhang, Xueying; Cao, Chunwei; Hambly, Catherine; Qin, Guosong; Yao, Jing; Song, Ruigao; Jia, Qitao; Wang, Xiao; Li, Yongshun; Zhang, Nan; Piao, Zhengyu; Ye, Rongcai; Speakman, John R; Wang, Hongmei; Zhou, Qi; Wang, Yanfang; Jin, Wanzhu; Zhao, Jianguo.

In: PNAS, Vol. 114, No. 45, 2017, p. 9474-9482.

Research output: Contribution to journalArticle

Zheng, Q, Lin, J, Huang, J, Zhang, H, Zhang, R, Zhang, X, Cao, C, Hambly, C, Qin, G, Yao, J, Song, R, Jia, Q, Wang, X, Li, Y, Zhang, N, Piao, Z, Ye, R, Speakman, JR, Wang, H, Zhou, Q, Wang, Y, Jin, W & Zhao, J 2017, 'Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity', PNAS, vol. 114, no. 45, pp. 9474-9482. https://doi.org/10.1073/pnas.1707853114
Zheng, Qiantao ; Lin, Jun ; Huang, Jiaojiao ; Zhang, Hongyong ; Zhang, Rui ; Zhang, Xueying ; Cao, Chunwei ; Hambly, Catherine ; Qin, Guosong ; Yao, Jing ; Song, Ruigao ; Jia, Qitao ; Wang, Xiao ; Li, Yongshun ; Zhang, Nan ; Piao, Zhengyu ; Ye, Rongcai ; Speakman, John R ; Wang, Hongmei ; Zhou, Qi ; Wang, Yanfang ; Jin, Wanzhu ; Zhao, Jianguo. / Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity. In: PNAS. 2017 ; Vol. 114, No. 45. pp. 9474-9482.
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abstract = "Uncoupling protein 1 (UCP1) is localized on the inner mitochondrial membrane and generates heat by uncoupling ATP synthesis from proton transit across the inner membrane. UCP1 is a key element of nonshivering thermogenesis and is most likely important in the regulation of body adiposity. Pigs (Artiodactyl family Suidae) lack a functional UCP1 gene, resulting in poor thermoregulation and susceptibility to cold, which is an economic and pig welfare issue owing to neonatal mortality. Pigs also have a tendency toward fat accumulation, which may be linked to their lack of UCP1, and thus influences the efficiency of pig production. Here, we report application of a CRISPR/Cas9-mediated, homologous recombination (HR)-independent approach to efficiently insert mouse adiponectin-UCP1 into the porcine endogenous UCP1 locus. The resultant UCP1 knock-in (KI) pigs showed an improved ability to maintain body temperature during acute cold exposure, but they did not have alterations in physical activity levels or total daily energy expenditure (DEE). Furthermore, ectopic UCP1 expression in white adipose tissue (WAT) dramatically decreased fat deposition by 4.89{\%} (P < 0.01), consequently increasing carcass lean percentage (CLP; P < 0.05). Mechanism studies indicated that the loss of fat upon UCP1 activation in WAT was linked to elevated lipolysis. UCP1 KI pigs are a potentially valuable resource for agricultural production through their combination of cold adaptation, which improves pig welfare and reduces economic losses, with reduced fat deposition and increased lean meat production.",
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author = "Qiantao Zheng and Jun Lin and Jiaojiao Huang and Hongyong Zhang and Rui Zhang and Xueying Zhang and Chunwei Cao and Catherine Hambly and Guosong Qin and Jing Yao and Ruigao Song and Qitao Jia and Xiao Wang and Yongshun Li and Nan Zhang and Zhengyu Piao and Rongcai Ye and Speakman, {John R} and Hongmei Wang and Qi Zhou and Yanfang Wang and Wanzhu Jin and Jianguo Zhao",
note = "Acknowledgments We thank members of the J.Z., W.J., and Y.W. laboratories for helpful discussions; P. Chai, S. Liu, H. Tang, and C. Wei (Institute of High Energy Physics at the Chinese Academy of Sciences and Beijing Engineering Research Center of Radiographic Techniques and Equipment) for their help with the PET scan experiments and analysis; L. Yang (Institute of Genetics and Developmental Biology at the Chinese Academy of Sciences) for help in TEM analysis; and Peter Thomson (University of Aberdeen) for technical assistance with isotope analysis. This study was supported by the National Transgenic Project of China (Grant 2016ZX08009003-006-007), the Strategic Priority Research Programs of the Chinese Academy of Sciences (Grants XDA08010304 and XDB13030000), the National Natural Science Foundation of China (Grants 81671274 and 31272440), and the National Program on Key Basic Research Project (973 Program; Grant 2015CB943100). Y.W. was supported by the Elite Youth Program of the Chinese Academy of Agricultural Sciences (ASTIP-IAS05). This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1707853114/-/DCSupplemental.",
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TY - JOUR

T1 - Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity

AU - Zheng, Qiantao

AU - Lin, Jun

AU - Huang, Jiaojiao

AU - Zhang, Hongyong

AU - Zhang, Rui

AU - Zhang, Xueying

AU - Cao, Chunwei

AU - Hambly, Catherine

AU - Qin, Guosong

AU - Yao, Jing

AU - Song, Ruigao

AU - Jia, Qitao

AU - Wang, Xiao

AU - Li, Yongshun

AU - Zhang, Nan

AU - Piao, Zhengyu

AU - Ye, Rongcai

AU - Speakman, John R

AU - Wang, Hongmei

AU - Zhou, Qi

AU - Wang, Yanfang

AU - Jin, Wanzhu

AU - Zhao, Jianguo

N1 - Acknowledgments We thank members of the J.Z., W.J., and Y.W. laboratories for helpful discussions; P. Chai, S. Liu, H. Tang, and C. Wei (Institute of High Energy Physics at the Chinese Academy of Sciences and Beijing Engineering Research Center of Radiographic Techniques and Equipment) for their help with the PET scan experiments and analysis; L. Yang (Institute of Genetics and Developmental Biology at the Chinese Academy of Sciences) for help in TEM analysis; and Peter Thomson (University of Aberdeen) for technical assistance with isotope analysis. This study was supported by the National Transgenic Project of China (Grant 2016ZX08009003-006-007), the Strategic Priority Research Programs of the Chinese Academy of Sciences (Grants XDA08010304 and XDB13030000), the National Natural Science Foundation of China (Grants 81671274 and 31272440), and the National Program on Key Basic Research Project (973 Program; Grant 2015CB943100). Y.W. was supported by the Elite Youth Program of the Chinese Academy of Agricultural Sciences (ASTIP-IAS05). This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1707853114/-/DCSupplemental.

PY - 2017

Y1 - 2017

N2 - Uncoupling protein 1 (UCP1) is localized on the inner mitochondrial membrane and generates heat by uncoupling ATP synthesis from proton transit across the inner membrane. UCP1 is a key element of nonshivering thermogenesis and is most likely important in the regulation of body adiposity. Pigs (Artiodactyl family Suidae) lack a functional UCP1 gene, resulting in poor thermoregulation and susceptibility to cold, which is an economic and pig welfare issue owing to neonatal mortality. Pigs also have a tendency toward fat accumulation, which may be linked to their lack of UCP1, and thus influences the efficiency of pig production. Here, we report application of a CRISPR/Cas9-mediated, homologous recombination (HR)-independent approach to efficiently insert mouse adiponectin-UCP1 into the porcine endogenous UCP1 locus. The resultant UCP1 knock-in (KI) pigs showed an improved ability to maintain body temperature during acute cold exposure, but they did not have alterations in physical activity levels or total daily energy expenditure (DEE). Furthermore, ectopic UCP1 expression in white adipose tissue (WAT) dramatically decreased fat deposition by 4.89% (P < 0.01), consequently increasing carcass lean percentage (CLP; P < 0.05). Mechanism studies indicated that the loss of fat upon UCP1 activation in WAT was linked to elevated lipolysis. UCP1 KI pigs are a potentially valuable resource for agricultural production through their combination of cold adaptation, which improves pig welfare and reduces economic losses, with reduced fat deposition and increased lean meat production.

AB - Uncoupling protein 1 (UCP1) is localized on the inner mitochondrial membrane and generates heat by uncoupling ATP synthesis from proton transit across the inner membrane. UCP1 is a key element of nonshivering thermogenesis and is most likely important in the regulation of body adiposity. Pigs (Artiodactyl family Suidae) lack a functional UCP1 gene, resulting in poor thermoregulation and susceptibility to cold, which is an economic and pig welfare issue owing to neonatal mortality. Pigs also have a tendency toward fat accumulation, which may be linked to their lack of UCP1, and thus influences the efficiency of pig production. Here, we report application of a CRISPR/Cas9-mediated, homologous recombination (HR)-independent approach to efficiently insert mouse adiponectin-UCP1 into the porcine endogenous UCP1 locus. The resultant UCP1 knock-in (KI) pigs showed an improved ability to maintain body temperature during acute cold exposure, but they did not have alterations in physical activity levels or total daily energy expenditure (DEE). Furthermore, ectopic UCP1 expression in white adipose tissue (WAT) dramatically decreased fat deposition by 4.89% (P < 0.01), consequently increasing carcass lean percentage (CLP; P < 0.05). Mechanism studies indicated that the loss of fat upon UCP1 activation in WAT was linked to elevated lipolysis. UCP1 KI pigs are a potentially valuable resource for agricultural production through their combination of cold adaptation, which improves pig welfare and reduces economic losses, with reduced fat deposition and increased lean meat production.

KW - Journal Article

KW - UCP1

KW - thermoregulation

KW - fat deposition

KW - pig

KW - CRISPR/Cas9

U2 - 10.1073/pnas.1707853114

DO - 10.1073/pnas.1707853114

M3 - Article

C2 - 29078316

VL - 114

SP - 9474

EP - 9482

JO - PNAS

JF - PNAS

SN - 0027-8424

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ER -