CRISPR/Cas9‐mediated sheep <i>MSTN</i> gene knockout and promote sSMSCs differentiation

Zhang, Yani; Wang, Yingjie; Yulin, Bi; Tang, Bao-Jun; Wang, Man; Zhang, Chen; Zhang, Wenhui; Jin, Jing; Li, Tingting; Zhao, Ruihua; Yu, Xudong; Zuo, Qisheng; Li, Bichun

doi:10.1002/jcb.27474

Cited by 43 publications

(26 citation statements)

References 17 publications

(32 reference statements)

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“…Moreover, disrupting the normal function of MSTN in sheep skeletal muscle satellite cells (sSMSCs) has been shown to promote sSMSC differentiation in both number and length. This study has also reported the generation of MSTN -disputed sheep using SCNT from CRISPR/Cas9 transfected ear fibroblasts (Zhang et al, 2018c). A further and more recent report, published by our team, described the application of CRISPR/Cas9-based base editors for the introduction of a point mutation within the suppressor of cytokine signaling 2 (SOCS2) gene in sheep (Zhou et al, 2019).…”

Section: Applications Of Crispr/cas9 In Sheep and Goatsmentioning

confidence: 89%

Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era

et al. 2019

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Sheep and goats are valuable livestock species that have been raised for their production of meat, milk, fiber, and other by-products. Due to their suitable size, short gestation period, and abundant secretion of milk, sheep and goats have become important model animals in agricultural, pharmaceutical, and biomedical research. Genome engineering has been widely applied to sheep and goat research. Pronuclear injection and somatic cell nuclear transfer represent the two primary procedures for the generation of genetically modified sheep and goats. Further assisted tools have emerged to enhance the efficiency of genetic modification and to simplify the generation of genetically modified founders. These tools include sperm-mediated gene transfer, viral vectors, RNA interference, recombinases, transposons, and endonucleases. Of these tools, the four classes of site-specific endonucleases (meganucleases, ZFNs, TALENs, and CRISPRs) have attracted wide attention due to their DNA double-strand break-inducing role, which enable desired DNA modifications based on the stimulation of native cellular DNA repair mechanisms. Currently, CRISPR systems dominate the field of genome editing. Gene-edited sheep and goats, generated using these tools, provide valuable models for investigations on gene functions, improving animal breeding, producing pharmaceuticals in milk, improving animal disease resistance, recapitulating human diseases, and providing hosts for the growth of human organs. In addition, more promising derivative tools of CRISPR systems have emerged such as base editors which enable the induction of single-base alterations without any requirements for homology-directed repair or DNA donor. These precise editors are helpful for revealing desirable phenotypes and correcting genetic diseases controlled by single bases. This review highlights the advances of genome engineering in sheep and goats over the past four decades with particular emphasis on the application of CRISPR/Cas9 systems.

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Section: Applications Of Crispr/cas9 In Sheep and Goatsmentioning

confidence: 89%

Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era

et al. 2019

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“…In broiler growth and development, the number of muscle fibers is mainly determined in the embryonic stage [ 34 ]. At the same time, myostatin ( MSTN ) is an important gene regulating embryonic muscle cells and adult muscle development, so MSTN is crucial for skeletal muscle growth and development [ 35 ]. Studies have shown that recombinant myostatin protein can inhibit C2C12 mouse myoblasts proliferation, DNA synthesis, and protein synthesis [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

MicroRNA-27b-3p Targets the Myostatin Gene to Regulate Myoblast Proliferation and Is Involved in Myoblast Differentiation

Zhang

et al. 2021

Cells

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microRNAs play an important role in the growth and development of chicken embryos, including the regulation of skeletal muscle genesis, myoblast proliferation, differentiation, and apoptosis. Our previous RNA-seq studies showed that microRNA-27b-3p (miR-27b-3p) might play an important role in regulating the proliferation and differentiation of chicken primary myoblasts (CPMs). However, the mechanism of miR-27b-3p regulating the proliferation and differentiation of CPMs is still unclear. In this study, the results showed that miR-27b-3p significantly promoted the proliferation of CPMs and inhibited the differentiation of CPMs. Then, myostatin (MSTN) was confirmed to be the target gene of miR-27b-3p by double luciferase reporter assay, RT-qPCR, and Western blot. By overexpressing and interfering with MSTN expression in CPMs, the results showed that overexpression of MSTN significantly inhibited the proliferation and differentiation of CPMs. In contrast, interference of MSTN expression had the opposite effect. This study showed that miR-27b-3p could promote the proliferation of CPMs by targeting MSTN. Interestingly, both miR-27b-3p and MSTN can inhibit the differentiation of CPMs. These results provide a theoretical basis for further understanding the function of miR-27b-3p in chicken and revealing its regulation mechanism on chicken muscle growth.

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“…Researchers generated marker-gene-free MSTN biallelic mutation cattle via ZFNs combined with SCNT technology, which exhibited the double-muscled phenotype with about half the level of MSTN protein with an intact C-terminal domain estimated by ELISA analysis [4] . MSTN-KO goats [24,52] and sheep [53] have also been generated via CRISPR/Cas9.…”

Section: Milk and Meatmentioning

confidence: 99%

The development and application of genome editing technology in ruminants: a review

Yuan¹,

Gao²,

Han³

et al. 2020

Front. Agr. Sci. Eng.

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Transgenic ruminants are a valuable resource for both animal breeding and biomedical research. The development of transgenic breeding is proceeding slowly, because it suffers from low efficiency of gene transfer and possible safety problems from uncontrolled random integration. However, new breeding methods combined with genome editing and somatic cell nuclear transfer or microinjection can offer an economic and efficient way to produce gene-edited ruminants, which can serve as bioreactors or have improved disease resistance, animal welfare and product quality. Recent advances in precise genome editing technologies, especially clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 nucleases, are enabling the systematic development of gene-edited ruminant production. This review covers the development of gene-edited ruminants, the particulars of site-specific engineered nucleases and the state of the art and new insights into practical applications and social acceptance of genome editing technology in ruminants. It is concluded that the production of gene-edited ruminants is feasible and through improvements in genome editing technology it is possible to help feed the world.

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CRISPR/Cas9‐mediated sheep MSTN gene knockout and promote sSMSCs differentiation

Cited by 43 publications

References 17 publications

Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era

Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era

MicroRNA-27b-3p Targets the Myostatin Gene to Regulate Myoblast Proliferation and Is Involved in Myoblast Differentiation

The development and application of genome editing technology in ruminants: a review

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