Generation of myostatin‐knockout chickens mediated by D10A‐Cas9 nickase

Kim, Gap-Don; Lee, Jeong Hyo; Song, Sumin; Kim, Seo Woo; Han, Ji Seon; Shin, Seung Pyo; Park, Byung‐Chul; Park, Tae Sub

doi:10.1096/fj.201903035r

Cited by 70 publications

(100 citation statements)

References 33 publications

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“…Interestingly, we found that myostatin (MSTN) was also one of top 20 down-regulated genes. MSTN, a member of transforming growth factor beta (TGFbeta) superfamily, has been reported as a negative regulator of muscle growth and development [29][30][31][32][33].…”

Section: Discussionmentioning

confidence: 99%

Insulin-degrading Enzyme Regulates the Proliferation and Apoptosis of Porcine Skeletal Muscle Stem Cells via Myostatin/MYOD Pathway

Wang

Guo

Zhang

et al. 2021

Preprint

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Background: Identifying the genes relevant for muscle development is pivotal to improve meat production and quality in pigs. Insulin-degrading enzyme (IDE), a thiol zinc-metalloendopeptidase, has been known to regulate the myogenic process of mouse and rat myoblast cell lines, while its myogenic role in pigs remained elusive. Therefore, the current study aimed to identify the effects of IDE on the proliferation and apoptosis of porcine skeletal muscle stem cells and underlying molecular mechanism.Results: We found in the present study that IDE was widely expressed in porcine tissues, including kidney, lung, spleen, liver, heart, and skeletal muscle. Then, to explore the effects of IDE on the proliferation and apoptosis of porcine skeletal muscle stem cells, we subjected the cells to siRNA-mediated knockdown of IDE expression, which resulted in promoted cell proliferation and reduced apoptosis. As one of key transcription factors in myogenesis, MYOD, its expression was also decreased with IDE knockdown. To further elucidate the underlying molecular mechanism, RNA sequencing was performed. Among transcripts perturbed by the IDE knockdown after, a down-regulated gene myostatin (MSTN) which is known as a negative regulator for muscle growth attracted our interest. Indeed, MSTN knockdown led to similar results as those of the IDE knockdown, with upregulation of cell cycle-related genes, downregulation of MYOD as well as apoptosis-related genes, and enhanced cell proliferation.Conclusion: Our findings suggest that IDE regulates the proliferation and apoptosis of porcine skeletal muscle stem cells through MSTN/MYOD pathway. Thus, we recruit IDE to the gene family of regulators for porcine skeletal muscle development, and propose IDE as an example of gene to prioritize in order to improve pork production.

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Section: Discussionmentioning

confidence: 99%

Insulin-degrading Enzyme Regulates the Proliferation and Apoptosis of Porcine Skeletal Muscle Stem Cells via Myostatin/MYOD Pathway

Wang

Guo

Zhang

et al. 2021

Preprint

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“…Studies to date have examined the gain and loss‐of‐function of MSTN in chicken embryos, the consequences for the muscle progenitor population, and the association between MSTN SNPs and broiler performance 39,40 . We did not further analyze the our MSTN genome‐modified chickens in detail, but recent studies showed that mutation of MSTN in chicken 41 and quail has an anti‐myogenic effect 42 ; moreover, spatial MSTN mutations introduced by adenoviral CRISPR‐Cas9 induce differential gene expression involved in cell differentiation, muscle growth and development, and energy metabolism in postnatal chicken 43 . Therefore, we expect that these animals will exhibit elevated protein synthesis capacity as well as improved muscle cell development.…”

Section: Discussionmentioning

confidence: 99%

Highly elevated base excision repair pathway in primordial germ cells causes low base editing activity in chickens

Lee

Han

et al. 2020

FASEB j.

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Base editing technology enables the generation of precisely genome-modified animal models. In this study, we applied base editing to chicken, an important livestock animal in the fields of agriculture, nutrition, and research through primordial germ cell (PGC)-mediated germline transmission. Using this approach, we successfully produced two genome-modified chicken lines harboring mutations in the genes encoding ovotransferrin (TF) and myostatin (MSTN); however, only 55.5% and 35.7% of genome-modified chickens had the desired base substitutions in TF and MSTN, respectively. To explain the low base-editing activity, we performed molecular analysis to compare DNA repair pathways between PGCs and the chicken fibroblast cell line DF-1. The results revealed that base excision repair (BER)-related genes were significantly elevated in PGCs relative to DF-1 cells. Subsequent functional studies confirmed that the editing activity could be regulated by modulating the expression of uracil N-glycosylase (UNG), an upstream gene of the BER pathway. Collectively, our findings indicate that the distinct DNA repair property of chicken PGCs causes low editing activity during genome modification, however, modulation of BER functions could promote the production of genome-modified organisms with the desired genotypes. K E Y W O R D S base editing, base excision repair, chicken, editing activity, primordial germ cell, uracil N-glycosylase 2 | LEE Et aL.

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“…In this regard, the myostatin (MSTN) gene gains huge attention, because MSTN is a negative regulator of muscle growth, and mutation in the MSTN gene resulted in increased muscle mass in mammals and fishes [92][93][94][95][96][97][98][99][100][101][102]. To investigate anti-myogenic function of MSTN in the avian species, MSTN knockout quail and chickens were generated by using the adenovirus-and PGC-mediated method, respectively [72,87]. Significant increase in breast and leg muscle in both MSTN knockout quail and chicken indicated that MSTN can be a potential selection marker for poultry lines with higher meat yield.…”

Section: Industrial Purposementioning

confidence: 99%

Current Approaches and Applications in Avian Genome Editing

Lee

Kim

Lee

2020

IJMS

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Advances in genome-editing technologies and sequencing of animal genomes enable researchers to generate genome-edited (GE) livestock as valuable animal models that benefit biological researches and biomedical and agricultural industries. As birds are an important species in biology and agriculture, their genome editing has gained significant interest and is mainly performed by using a primordial germ cell (PGC)-mediated method because pronuclear injection is not practical in the avian species. In this method, PGCs can be isolated, cultured, genetically edited in vitro, and injected into a recipient embryo to produce GE offspring. Recently, a couple of GE quail have been generated by using the newly developed adenovirus-mediated method. Without technically required in vitro procedures of the PGC-mediated method, direct injection of adenovirus into the avian blastoderm in the freshly laid eggs resulted in the production of germ-line chimera and GE offspring. As more approaches are available in avian genome editing, avian research in various fields will progress rapidly. In this review, we describe the development of avian genome editing and scientific and industrial applications of GE avian species.

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Generation of myostatin‐knockout chickens mediated by D10A‐Cas9 nickase

Cited by 70 publications

References 33 publications

Insulin-degrading Enzyme Regulates the Proliferation and Apoptosis of Porcine Skeletal Muscle Stem Cells via Myostatin/MYOD Pathway

Insulin-degrading Enzyme Regulates the Proliferation and Apoptosis of Porcine Skeletal Muscle Stem Cells via Myostatin/MYOD Pathway

Highly elevated base excision repair pathway in primordial germ cells causes low base editing activity in chickens

Current Approaches and Applications in Avian Genome Editing

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