One-step generation of knockout pigs by zygote injection of CRISPR/Cas system

Hai, Tang; Teng, Fei; Guo, Runfa; Li, Wei; Zhou, Qi

doi:10.1038/cr.2014.11

Cited by 414 publications

(337 citation statements)

References 15 publications

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“…MSTN dysfunction resulted in dramatic increase of animal muscle mass due to hypertrophy and hyperplasia of muscle fibers [24][25][26][27]. Inhibition of MSTN expression by gene knockout could promote the muscle growth and meat production of livestock animals.…”

Section: Discussionmentioning

confidence: 99%

Double-Muscled Phenotype in Mutant Sheep Directed by the CRISPRCas9 System

Wu¹,

Du²,

Liu³

et al. 2018

Clon Transgen

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Myostatin (MSTN) is a well-known negative regulator of muscle growth. The double-muscled sheep caused by natural loss-of-function mutations of MSTN have very strong skeletal muscle. In this study, our results demonstrate the successful generation of MSTN mutant sheep via specific targeting of an exon 1 site using Cas9 technology. The MSTN-knockout sheep in our study had increased muscle significantly just like double-muscled phenotype. Our study suggests that the direct injection of Cas9: sgRNA into zygotes could be widely used to create gene knockouts in large domestic animals. Notably, on the basis of our findings, sheep can be added to the growing list of species for which genome editing is now practical. The generation of MSTN mutant sheep has implications for the genetic improvement of local sheep varieties, and also for the usage of sheep as a model for large animal medical research.

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Section: Discussionmentioning

confidence: 99%

Double-Muscled Phenotype in Mutant Sheep Directed by the CRISPRCas9 System

Wu¹,

Du²,

Liu³

et al. 2018

Clon Transgen

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“…With the platform of the CRISPR-Cas9 system, Dr. Zhou and his team achieved simultaneous mutations of multiple genes in rats , generated live vWFknockout pig models using a one-step method (Hai et al, 2014), and produced the first live p53 homozygous mutant monkey (Wan et al, 2015). These studies provide a new approach for rapid generation of genetically modified animal models of human diseases, and will promote research on pathogenesis and treatment of diseases.…”

Section: Generating Important Animal and Cell Models For Stem Cell Rementioning

confidence: 99%

Profile of Qi Zhou

2016

Sci. China Life Sci.

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Zhou has focused on stem cell research and has led a group carrying out a series of projects in the field of stem cell biology, with emphasis on studying the mechanisms of cell reprogramming and pluripotency of embryonic stem cells. He has also promoted the translation of stem cells to clinical applications. During the past years, Dr. Zhou has made many outstanding achievements. Developing novel methods for somatic cell reprogramming

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“…Moreover, after the generation of the first transgenic piglet by sperm injection, ICSI gained importance as a new tool for inducing genetic modifications in farm animals (Kurome et al 2006, Garcia-Vazquez et al 2010. However, the advent of CRISPR-Cas9 system for genetic engineering lead to the replacement of ICSI by regular or IVF zygotes for the generation of genetically modified animals (Hai et al 2014, Whitworth et al 2014, Proudfoot et al 2015, Wang et al 2015, Bevacqua et al 2016.…”

Section: General Overviewmentioning

confidence: 99%

“…In conclusion, ICSI-MGT in farm animal has only produced repeatable results in pigs, wherein several transgenic offspring have resulted. However, interest in producing transgenic animals by ICSI-MGT has decreased with the advent of new tools of gene edition (CRISPR-Cas9 and TALEN system), which are technically simpler (Hai et al 2014, Whitworth et al 2014, Proudfoot et al 2015, Wang et al 2015, Bevacqua et al 2016). …”

Section: Icsi-mediated Gene Transfer (Icsi-mgt)mentioning

confidence: 99%

Intracytoplasmic sperm injection in domestic and wild mammals

Salamone¹,

Canel²,

Rodríguez³

2017

Reproduction

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Intracytoplasmic sperm injection (ICSI) has become a useful technique for clinical applications in the horse-breeding industry. However, both ICSI blastocyst and offspring production continues to be limited for most farm and wild species. This article reviews technical differences of ICSI performance among species, possible biological and methodological reasons for the variable efficiency and potential strategies to improve the outcomes. One of the major applications of ICSI in animal production is the reproduction of high-value specimens. Unfortunately, some domestic species like the bovine show low rates of pronuclei formation after sperm injection, which led to the development of various artificial activation protocols and sperm pre-treatments that are discussed in this article. The impact of ICSI technique on equine breeding programs is considered in detail, since in contrast to other species, its use for elite horse reproduction has increased in recent years. ICSI has also been used to produce genetically modified animals; however, despite numerous attempts in several domestic species, only transgenic pigs have been consistently produced. Finally, the ICSI is a promising tool for genetic rescue of endangered and wild species. In conclusion, while ICSI has become a consistent ART for some species, it needs further development for others. The low results obtained for some domestic species, the high training needed and the equipment required have limited this technique to the production of elite specimens or for research purposes.Reproduction (2017) 154 F111-F124

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One-step generation of knockout pigs by zygote injection of CRISPR/Cas system

Cited by 414 publications

References 15 publications

Double-Muscled Phenotype in Mutant Sheep Directed by the CRISPRCas9 System

Double-Muscled Phenotype in Mutant Sheep Directed by the CRISPRCas9 System

Profile of Qi Zhou

Intracytoplasmic sperm injection in domestic and wild mammals

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