Open Access
PLoS ONE, volume 12, issue 10, pages e0186056
Generation of beta-lactoglobulin knock-out goats using CRISPR/Cas9
Publication type: Journal Article
Publication date: 2017-10-10
PubMed ID:
29016691
Multidisciplinary
Abstract
Goat’s milk, considered a substitute for cow’s milk, has a high nutritional value. However, goat’s milk contains various allergens, predominantly β-lactoglobulin (BLG). In this study, we employed the CRISPR/Cas9 system to target the BLG locus in goat fibroblasts for sgRNA optimization and generate BLG knock-out goats through co-injection of Cas9 mRNA and small guide RNAs (sgRNAs) into goat embryos at the one-cell stage. We firstly tested sgRNA editing efficiencies in goat fibroblast cells, and approximately 8.00%–9.09% of the cells were modified in single sgRNA-guided targeting experiment. Among the kids, the genome-targeting efficiencies of single sgRNA were 12.5% (10 ng/μL sg1) and 0% (10 ng/μL sg2) and efficiencies of dual sgRNAs were 25.0% (25 ng/μL sg2+sg3 group) and 28.6% (50 ng/μL sg2+sg3 group). Relative expression of BLG in BLG knock-out goat mammary glands significantly (p < 0.01) decreased as well as other milk protein coding genes, such as CSN1S1, CSN1S2, CSN2, CSN3 and LALBA (p < 0.05). As expected, BLG protein had been abolished in the milk of the BLG knock-out goat. In addition, most of the targeted kids were chimeric (3/4), and their various body tissues were edited simultaneously. Our study thus provides a basis for optimizing the quality of goat milk, which can be applied to biomedical and agricultural research.
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- We do not take into account publications that without a DOI.
- Statistics recalculated only for publications connected to researchers, organizations and labs registered on the platform.
- Statistics recalculated weekly.
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Zhou W. et al. Generation of beta-lactoglobulin knock-out goats using CRISPR/Cas9 // PLoS ONE. 2017. Vol. 12. No. 10. p. e0186056.
GOST all authors (up to 50)
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Zhou W., Wan Y., Guo R., Deng M., Deng K., Wang Z., Zhang Y., Wang F. Generation of beta-lactoglobulin knock-out goats using CRISPR/Cas9 // PLoS ONE. 2017. Vol. 12. No. 10. p. e0186056.
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TY - JOUR
DO - 10.1371/journal.pone.0186056
UR - https://doi.org/10.1371%2Fjournal.pone.0186056
TI - Generation of beta-lactoglobulin knock-out goats using CRISPR/Cas9
T2 - PLoS ONE
AU - Zhou, Wenjun
AU - Wan, Yongjie
AU - Guo, Rihong
AU - Deng, Mingtian
AU - Deng, Kaiping
AU - Wang, Zhen
AU - Zhang, Yanli
AU - Wang, Feng
PY - 2017
DA - 2017/10/10 00:00:00
PB - Public Library of Science (PLoS)
SP - e0186056
IS - 10
VL - 12
PMID - 29016691
SN - 1932-6203
ER -
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@article{2017_Zhou,
author = {Wenjun Zhou and Yongjie Wan and Rihong Guo and Mingtian Deng and Kaiping Deng and Zhen Wang and Yanli Zhang and Feng Wang},
title = {Generation of beta-lactoglobulin knock-out goats using CRISPR/Cas9},
journal = {PLoS ONE},
year = {2017},
volume = {12},
publisher = {Public Library of Science (PLoS)},
month = {oct},
url = {https://doi.org/10.1371%2Fjournal.pone.0186056},
number = {10},
pages = {e0186056},
doi = {10.1371/journal.pone.0186056}
}
Cite this
MLA
Copy
Zhou, Wenjun, et al. “Generation of beta-lactoglobulin knock-out goats using CRISPR/Cas9.” PLoS ONE, vol. 12, no. 10, Oct. 2017, p. e0186056. https://doi.org/10.1371%2Fjournal.pone.0186056.