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RNA-seq reveals transcriptome changes in goats following myostatin gene knockout
Lamei Wang1, Bei Cai1, Shiwei Zhou1
1College of Animal Science and Technology, Northwest A&F University, Yangling, China.
Plos One
|December 12, 2017
Summary
Myostatin (MSTN) gene editing in Shaanbei Cashmere goats using CRISPR/Cas9 revealed significant changes in fatty acid metabolism. MSTN knockout goats show altered gene expression related to lipid metabolism, offering insights into goat genomics.
Area of Science:
- Animal Genomics
- Molecular Biology
- Biochemistry
Background:
- Myostatin (MSTN) is a key negative regulator of skeletal muscle mass in mammals.
- Mutations in the MSTN gene are associated with the double-muscling trait.
- Understanding MSTN's role in goats is crucial for livestock improvement.
Purpose of the Study:
- To generate Myostatin (MSTN) knockout Shaanbei Cashmere goats using CRISPR/Cas9.
- To analyze the transcriptome profiles of MSTN-edited goats compared to wild-type and FGF5 knockout goats.
- To identify genes and pathways regulated by MSTN in goat skeletal muscle.
Main Methods:
- CRISPR/Cas9 gene editing was employed to create MSTN knockout goats.
- RNA sequencing (Illumina HiSeq 2000) was performed on muscle tissues.
- Transcriptome data were mapped to the Capra hircus reference genome using TopHat.
- Differentially expressed genes (DEGs) were identified and analyzed between groups.
- Quantitative real-time PCR (qRT-PCR) was used for validation.
Main Results:
- Significant numbers of differentially expressed genes (DEGs) were identified between wild-type, FGF5 knockout, and MSTN/FGF5 double knockout groups.
- MSTN knockout significantly altered gene expression, particularly in pathways related to fatty acid metabolism and unsaturated fatty acid biosynthesis.
- Key genes like stearoyl-CoA dehydrogenase and fatty acid synthase showed altered expression, suggesting MSTN's role in lipid metabolism.
- qRT-PCR results validated the RNA sequencing findings for selected DEGs.
Conclusions:
- MSTN plays a significant role in regulating lipid metabolism in goat skeletal muscle.
- MSTN knockout goats exhibit a unique transcriptome profile with substantial changes in fatty acid metabolism pathways.
- This study provides valuable genomic insights into MSTN function and its potential as a target for improving goat traits.
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