TY - JOUR
T1 - Transcriptional control of a novel long noncoding RNA Mymsl in smooth muscle cells by a single Cis-element and its initial functional characterization in vessels
AU - Choi, Mihyun
AU - Lu, Yao Wei
AU - Zhao, Jinjing
AU - Wu, Mingfu
AU - Zhang, Wei
AU - Long, Xiaochun
N1 - Funding Information:
We thank Dr. Harold Singer for critical comments on the manuscript, Mrs. Diane Singer for providing us the primary mouse VSMC cultures, and the University of Rochester Genomics Research Center for performing the RNA-seq experiments. This work is supported by National Institutes of Health R01HL122686 and R01HL139794 and Albany Medical College faculty startup funding to X.L; American Heart Association Scientist Development Grant 13SDG16920099 and National Institutes of Health R01HL121700 to M.W; American Heart Association Career Development Award (CDA) 18CDA34110319 to W.Z. M. Choi and X. Long designed and performed the research, analyzed the data, and wrote the paper. Y.W. Lu and J. Zhao performed experiments and analyzed the data. W. Zhang and M. Wu designed the research and edited the paper. The authors declare no conflicts of interest.
Funding Information:
This work is supported by National Institutes of Health R01HL122686 and R01HL139794 and Albany Medical College faculty startup funding to X.L; American Heart Association Scientist Development Grant 13SDG16920099 and National Institutes of Health R01HL121700 to M.W; American Heart Association Career Development Award (CDA) 18CDA34110319 to W.Z.
Publisher Copyright:
© 2019 Elsevier Ltd
PY - 2020/1
Y1 - 2020/1
N2 - Differentiated vascular smooth muscle cells (VSMCs) are crucial in maintaining vascular homeostasis. While the coding transcriptome of the differentiated VSMC phenotype has been defined, we know little about its noncoding signature. Herein, we identified a Myocardin-induced muscle specific long noncoding RNA (lncRNA) (Mymsl) downregulated upon VSMC phenotypic modulation. We demonstrated an essential role of a proximal consensus CArG element in response to MYOCD/SRF in vitro. To validate the in vivo role of this CArG element, we generated CArG mutant mice via CRISPR-Cas9 genome editing. While the CArG mutation had no impact on the expression of surrounding genes, it abolished Mymsl expression in SMCs, but not skeletal and cardiac muscle. Chromatin immunoprecipitation assays (ChIPs) showed decreased SRF binding to CArG region in mutants whereas the enrichment of H3K79Me2 remained the same. RNA-seq analysis showed a downregulation of matrix genes in aortas from Mymsl knockout mice, which was further validated in injured carotid arteries. Our study defined the transcriptional control of a novel lncRNA in SMCs via a single transcription factor binding site, which may offer a new strategy for generating SMC-specific knockout mouse models. We also provided in vivo evidence supporting the potential importance of Mymsl in vascular pathophysiology.
AB - Differentiated vascular smooth muscle cells (VSMCs) are crucial in maintaining vascular homeostasis. While the coding transcriptome of the differentiated VSMC phenotype has been defined, we know little about its noncoding signature. Herein, we identified a Myocardin-induced muscle specific long noncoding RNA (lncRNA) (Mymsl) downregulated upon VSMC phenotypic modulation. We demonstrated an essential role of a proximal consensus CArG element in response to MYOCD/SRF in vitro. To validate the in vivo role of this CArG element, we generated CArG mutant mice via CRISPR-Cas9 genome editing. While the CArG mutation had no impact on the expression of surrounding genes, it abolished Mymsl expression in SMCs, but not skeletal and cardiac muscle. Chromatin immunoprecipitation assays (ChIPs) showed decreased SRF binding to CArG region in mutants whereas the enrichment of H3K79Me2 remained the same. RNA-seq analysis showed a downregulation of matrix genes in aortas from Mymsl knockout mice, which was further validated in injured carotid arteries. Our study defined the transcriptional control of a novel lncRNA in SMCs via a single transcription factor binding site, which may offer a new strategy for generating SMC-specific knockout mouse models. We also provided in vivo evidence supporting the potential importance of Mymsl in vascular pathophysiology.
KW - CRISPR genome editing
KW - Long noncoding RNA
KW - Transcription factor
KW - Vascular smooth muscle
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UR - https://www.mendeley.com/catalogue/8ee3e0a4-3571-377f-b40c-b52cc25b33d6/
U2 - 10.1016/j.yjmcc.2019.11.148
DO - 10.1016/j.yjmcc.2019.11.148
M3 - Article
C2 - 31751568
AN - SCOPUS:85075833010
SN - 0022-2828
VL - 138
SP - 147
EP - 157
JO - Journal of molecular and cellular cardiology
JF - Journal of molecular and cellular cardiology
ER -