Journal ArticleDOI
RAPID COMMUNICATION: Generation of FGF5 knockout sheep via the CRISPR/Cas9 system
R. Hu,Z. Y. Fan,B. Y. Wang,Shou-Long Deng,X. S. Zhang,J. L. Zhang,Hongbing Han,Zhengxing Lian +7 more
TLDR
This study generated 3 knockout sheep via the 1-step clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system and discovered that wool length in founders was significantly longer than in wild types.Abstract:
Sheep are an important source of fiber production. Fibroblast growth factor 5 (FGF5) is a dominant inhibitor of length of the anagen phase of the hair cycle. Knockout or silencing of the gene results in a wooly coat in mice, donkeys, dogs, and rabbits. In sheep breeding, wool length is one of the most important wool quality traits. However, traditional breeding cannot accurately and efficiently mediate an advanced genotype into the sheep genome. In this study, we generated 3 knockout sheep via the 1-step clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system. Sequencing analysis confirmed that mutations in the gene existed in all germ lines of 3 founders: besides the intact sequence, 3 kinds of deletions in the gene (including 5, 13, and 33 bp) were detected. The changes in the primary and senior structure of the FGF5 protein due to the 3 deletions in founders suggested that the FGF5 protein was dysfunctional. In addition, the expression level of intact mRNA in heterozygous individuals decreased compared with the wild types ( < 0.01). Functionally, we discovered that wool length in founders was significantly longer than in wild types ( < 0.05). Collectively, the knockout sheep with the longer wool length phenotype will provide an efficient way for fast genetic improvement of sheep breeding and promote the development of wool industry.read more
Citations
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Journal ArticleDOI
Improving pregnancy outcomes in humans through studies in sheep
Janna L. Morrison,Mary J. Berry,Kimberley J. Botting,Jack R. T. Darby,Martin G. Frasch,Kathryn L. Gatford,Dino A. Giussani,Clint Gray,Richard Harding,Emilio A. Herrera,Matthew W. Kemp,Mitchell C. Lock,I. Caroline McMillen,Timothy J. M. Moss,Gabrielle C. Musk,Mark H. Oliver,Timothy R. H. Regnault,Claire T. Roberts,Jia Yin Soo,Ross L. Tellam +19 more
TL;DR: The aim of this review is to highlight the advances in perinatal human medicine that have been achieved following translation of research using the pregnant sheep and fetus.
Journal ArticleDOI
Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era.
Peter Kalds,Shiwei Zhou,Bei Cai,Jiao Liu,Ying Wang,Bjoern Petersen,Tad S. Sonstegard,Xiaolong Wang,Yulin Chen +8 more
TL;DR: Gene-edited sheep and goats, generated using these tools, provide valuable models for investigations on gene functions, improving animal breeding, producing pharmaceuticals in milk, improvingAnimal disease resistance, recapitulating human diseases, and providing hosts for the growth of human organs.
Journal ArticleDOI
CRISPR/Cas9 microinjection in oocytes disables pancreas development in sheep
Marcela Vilarino,Sheikh Tamir Rashid,Sheikh Tamir Rashid,Fabian P. Suchy,Bret R. McNabb,Talitha van der Meulen,Eli J. Fine,Syed Daniyal Ahsan,Syed Daniyal Ahsan,Nurlybek Mursaliyev,Vittorio Sebastiano,Santiago S. Diab,Mark O. Huising,Hiromitsu Nakauchi,Pablo J. Ross +14 more
TL;DR: The creation of pancreatogenesis-disabled sheep by oocyte microinjection of CRISPR/Cas9 targeting PDX1, a critical gene for pancreas development is reported, which provides the basis for the production of gene-edited sheep as a host for interspecies organ generation.
Journal ArticleDOI
Genome editing approaches to augment livestock breeding programs.
TL;DR: Genome editing could complement traditional livestock genetic improvement programs by fixing favorable alleles, introducing useful genetic variation and accelerating the rate of genetic gain.
Journal ArticleDOI
CRISPR/Cas9-mediated loss of FGF5 function increases wool staple length in sheep.
Wen Rong Li,Chen Xi Liu,Xue Mei Zhang,Lei Chen,Xin Rong Peng,San Gang He,Jia Peng Lin,Bin Han,Li Qin Wang,Jun Cheng Huang,Ming Jun Liu +10 more
TL;DR: It is suggested that CRISPR/Cas9‐mediated loss of FGF5 activity could promote wool growth and, consequently, increase wool length and yield.
References
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TL;DR: The CRISPR/Cas system allows the one-step generation of animals carrying mutations in multiple genes, an approach that will greatly accelerate the in vivo study of functionally redundant genes and of epistatic gene interactions.
Development and Applications of CRISPR-Cas9 for Genome Engineering
TL;DR: The development and applications of Cas9 are described for a variety of research or translational applications while highlighting challenges as well as future directions.
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