Systematic Identification and Comparison of the Expressed Profiles of lncRNAs, miRNAs, circRNAs, and mRNAs with Associated Co-Expression Networks in Pigs with Low and High Intramuscular Fat
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Ethics Statement
2.2. Animal and Sample Preparation
2.3. Construction and Sequencing of cDNA Libraries
2.4. Data Mapping and Transcriptome Assembly
2.5. Identification of lncRNAs, miRNAs, and circRNA
2.6. Differentially Expressed RNA Analysis
2.7. Prediction of the Potential Target Genes of DE lncRNAs, miRNAs, and circRNAs
2.8. Gene Ontology Enrichment and KEGG Pathway Analyses
2.9. Co-Construction of Gene Expression Networks
2.10. Association Analysis between QTL Sites and the Locations of Differentially Expressed RNA
2.11. Validation of the RNA Sequencing Results Using qRT-PCR
2.12. Statistical Analyses
3. Results
3.1. Overview of RNA Sequencing
3.2. Differential Expression Profiles ofmRNAs lncRNAs, miRNAs, and circRNAs
3.3. Prediction of the Potential Target Genes (PTGs) of DE lncRNAs, circRNAs, and miRNAs
3.4. GO and KEGG Analysis of the DERs
3.5. Functional Analysis of the DE PTGs
3.6. Overlapping Analysis between QTL Sites and the Location of DE RNAs
3.7. Expression Regulation Analysis of DE lncRNAs, miRNAs, and circRNAs, and Their DE PTGs
3.8. RNA Sequencing Results Validation Using qRT-PCR
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
ΔΔCt | delta delta cycle threshold |
ACAA2 | Acetyl-CoA Acyltransferase 2 |
ACOX2 | Acyl-CoA oxidase 2 |
CCL4 | C-C Motif Chemokine Ligand 4 |
CCL10 | C-C Motif Chemokine Ligand 10 |
CIB2 | calcium and integrin binding family member 2 |
ceRNA | competing endogenous RNAs |
circRNAs | circular RNAs |
CXCL10 | C-X-C motif chemokine ligand 10 |
CXCL16 | C-X-C motif chemokine ligand 16 |
DE | Differential expression |
DERs | differentially expressed RNAs |
FPKM | Fragments per kilobase of exon model per million mapped fragments |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
GO | Gene ontology |
GPAT | Glycerol-3-phosphate ethyltransferase |
GPNMB | glycoprotein nmb. |
GWAS | genome-wide association study |
HUS1 | HUS1 Checkpoint Clamp Component |
IMF | Intramuscular fat |
KEGG | Kyoto Encyclopedia of Genes and Genomes |
KCNRG | Potassium Channel Regulator |
lncRNAs | long non-coding RNAs |
LDM | longissimus dorsi muscle |
mRNAs | message RNA |
miRNAs | microRNAs |
miRNAs | microRNAs |
MYH7B | myosin heavy chain 7B |
Novel | novel gene |
NPPC | National Pork Producers Council |
PTGs | potential target genes |
PTPMT1 | protein tyrosine phosphatase mitochondrial 1 |
qRT-PCR | Reverse transcription quantitative polymerase chain reaction |
QTLs | quantitative trait loci |
RABL2B | RAB, member of ras oncogene family like 2b |
RDH16 | retinol dehydrogenase 16 |
rRNAs | mitochondrial ribosomal RNAs |
SFRP4 | secreted frizzled-related protein 4 |
sRNA | small RNA |
snRNAs | small nuclear RNAs |
snoRNA | small nucleolar RNA |
SPP1 | secreted phosphoprotein 1 |
TGFB3 | transforming growth factor beta 3 |
THBS4 | thrombospondin 4 |
tRNA | transfer RNA |
MAPK10 | mitogen-activated protein kinase 10 |
JAK1 | janus kinase 1 |
STAT1 | signal transducer and activator of transcription 1 |
TYK2 | tyrosine kinase 2 |
IRF9 | interferon regulatory factor 9 |
FADD | fas associated via death domain |
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Sample | Carcass Weight (kg) | IMF (%) | Group |
---|---|---|---|
H1 | 82.6 | 4.07 | High IMF |
H2 | 113 | 4.40 | High IMF |
H3 | 113 | 4.56 | High IMF |
H4 | 97.4 | 4.98 | High IMF |
H5 | 96.8 | 5.43 | High IMF |
L1 | 92 | 1.05 | Low IMF |
L2 | 113 | 1.18 | Low IMF |
L3 | 125.6 | 1.28 | Low IMF |
L4 | 89.4 | 1.57 | Low IMF |
L5 | 94.4 | 1.60 | Low IMF |
RNA | Regulated | log2FC | p-Value | Type |
---|---|---|---|---|
MSTRG.19330.20 | up | 6.573623 | 5.41 × 10−62 | lncRNAs |
MSTRG.40179.2 | up | 3.10731 | 1.32 × 10−10 | |
MSTRG.44176.8 | up | 1.908359 | 2.75 × 10−5 | |
MSTRG.39829.10 | up | 1.710734 | 4.15 × 10−4 | |
MSTRG.38601.10 | up | 1.476951 | 1.54 × 10−6 | |
MSTRG.29140.1 | down | −1.94114 | 2.37 × 10−5 | |
MSTRG.25219.1 | down | −1.99459 | 2.95 × 10−5 | |
MSTRG.9199.1 | down | −2.01282 | 4.72 × 10−6 | |
MSTRG.5761.2 | down | −2.50226 | 3.96 × 10−8 | |
MSTRG.44725.16 | down | −2.71452 | 1.45 × 10−10 | |
novel_miR_118 | up | 1.459799 | 1.07 × 10−2 | miRNAs |
ssc-miR-208b | up | 1.310524 | 7.14 × 10−3 | |
novel_miR_398 | up | 1.297012 | 3.44 × 10−2 | |
novel_miR_278 | up | 1.272563 | 3.34 × 10−2 | |
ssc-miR-190b | up | 1.198435 | 1.12 × 10−2 | |
ssc-miR-499-5p | up | 1.185263 | 1.01 × 10−2 | |
novel_miR_185 | down | −1.54969 | 1.09 × 10−2 | |
novel_miR_45 | down | −1.74161 | 3.76 × 10−3 | |
novel_miR_476 | down | −1.78496 | 3.59 × 10−3 | |
novel_miR_45 | down | −1.74161 | 3.76 × 10−3 | |
novel_miR_476 | down | −1.78496 | 3.59 × 10−3 | |
12:39408156|39428231 | up | 9.005799 | 1.98 × 10−6 | circRNAs |
14:71348983|71349948 | up | 6.965969 | 4.78 × 10−4 | |
3:44121881|44122061 | up | 6.819521 | 3.29 × 10−4 | |
9:125732918|125735258 | up | 6.586107 | 1.03 × 10−3 | |
13:71794794|71797638 | up | 6.346968 | 1.25 × 10−3 | |
1:108385212|108386218 | down | −5.96632 | 1.61 × 10−3 | |
12:59320434|59323398 | down | −6.16306 | 1.81 × 10−3 | |
7:68514625|68532510 | down | −6.37264 | 1.20 × 10−3 | |
9:66405629|66409132 | down | −6.57113 | 8.32 × 10−4 | |
4:50433434|50447885 | down | −6.83942 | 4.52 × 10−4 | |
RDH16 | up | 0.993121 | 1.20 × 10−4 | mRNAs |
ENSSSCG00000045560 | up | 0.928328 | 2.29 × 10−4 | |
KCNRG | up | 0.856391 | 6.58 × 10−4 | |
ENSSSCG00000045892 | up | 0.844323 | 8.67 × 10−4 | |
RABL2B | up | 0.831953 | 1.16 × 10−3 | |
SPP1 | down | −0.92796 | 2.69 × 10−4 | |
CIB2 | down | −0.93599 | 8.18 × 10−5 | |
PTPMT1 | down | −0.96233 | 5.00 × 10−5 | |
MYH7B | down | −0.99074 | 1.05 × 10−5 | |
GPNMB | down | −1.15542 | 1.07 × 10−5 |
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Cheng, F.; Liang, J.; Yang, L.; Lan, G.; Wang, L.; Wang, L. Systematic Identification and Comparison of the Expressed Profiles of lncRNAs, miRNAs, circRNAs, and mRNAs with Associated Co-Expression Networks in Pigs with Low and High Intramuscular Fat. Animals 2021, 11, 3212. https://doi.org/10.3390/ani11113212
Cheng F, Liang J, Yang L, Lan G, Wang L, Wang L. Systematic Identification and Comparison of the Expressed Profiles of lncRNAs, miRNAs, circRNAs, and mRNAs with Associated Co-Expression Networks in Pigs with Low and High Intramuscular Fat. Animals. 2021; 11(11):3212. https://doi.org/10.3390/ani11113212
Chicago/Turabian StyleCheng, Feng, Jing Liang, Liyu Yang, Ganqiu Lan, Lixian Wang, and Ligang Wang. 2021. "Systematic Identification and Comparison of the Expressed Profiles of lncRNAs, miRNAs, circRNAs, and mRNAs with Associated Co-Expression Networks in Pigs with Low and High Intramuscular Fat" Animals 11, no. 11: 3212. https://doi.org/10.3390/ani11113212