Background: MicroRNAs (miRNAs) are endogenously expressed small noncoding RNAs with a length of about 22 nucleotides and play critical roles in the regulation of posttranscriptional gene expression. miR-487b-3p has been recently identified as a key regulator in skeletal muscle growth and development. However, the function of miR-487b-3p on goat skeletal muscle remains to be investigated.
Results: In this study, we found that miR-487b-3p was widespread in goat different tissues, with a significantly higher expression in muscle, especially in lamb. The results demonstrated that the expression of miR-487b-3p was gradually up-regulated during myoblast proliferation. Then, miR-487b-3p knockout primary goat myoblast cells clones were obtained by using CRISPR/Cas9 system and our RPG surrogate reporter-based screening. Further investigation uncovered that the knockout of miR-487b-3p significantly accelerated primary goat myoblast proliferation, which was accompanied by up-regulation of cell cycle-related genes, such as PCNA, Cyclin E and CDK2. What’s more, we found that miR-487b-3p targets directly the 3'UTR of insulin receptor substrate 1 (IRS1) gene and IRS1 knockdown by siRNA was able to down-regulate the expression of the cell cycle-related genes.
Conclusions: Collectively, these founding demonstrated miR-487b-3p as a potent inhibitor of cell proliferation which functions by targeting IRS1 gene in primary goat myoblast.
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Posted 12 Mar, 2021
Posted 12 Mar, 2021
Background: MicroRNAs (miRNAs) are endogenously expressed small noncoding RNAs with a length of about 22 nucleotides and play critical roles in the regulation of posttranscriptional gene expression. miR-487b-3p has been recently identified as a key regulator in skeletal muscle growth and development. However, the function of miR-487b-3p on goat skeletal muscle remains to be investigated.
Results: In this study, we found that miR-487b-3p was widespread in goat different tissues, with a significantly higher expression in muscle, especially in lamb. The results demonstrated that the expression of miR-487b-3p was gradually up-regulated during myoblast proliferation. Then, miR-487b-3p knockout primary goat myoblast cells clones were obtained by using CRISPR/Cas9 system and our RPG surrogate reporter-based screening. Further investigation uncovered that the knockout of miR-487b-3p significantly accelerated primary goat myoblast proliferation, which was accompanied by up-regulation of cell cycle-related genes, such as PCNA, Cyclin E and CDK2. What’s more, we found that miR-487b-3p targets directly the 3'UTR of insulin receptor substrate 1 (IRS1) gene and IRS1 knockdown by siRNA was able to down-regulate the expression of the cell cycle-related genes.
Conclusions: Collectively, these founding demonstrated miR-487b-3p as a potent inhibitor of cell proliferation which functions by targeting IRS1 gene in primary goat myoblast.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
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