Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein
Open accessAbstract
Comprehensive studies have shown that DNA methylation plays vital roles in both loss of pluripotency and governance of the transcriptome during embryogenesis and subsequent developmental processes. Aberrant DNA methylation patterns have been widely observed in tumorigenesis, ageing and neurodegenerative diseases, highlighting the importance of a systematic understanding of DNA methylation and the dynamic changes of methylomes during disease onset and progression. Here we describe a facile and convenient approach for efficient targeted DNA methylation by fusing inactive Cas9 (dCas9) with an engineered prokaryotic DNA methyltransferase MQ1. Our study presents a rapid and efficient strategy to achieve locus-specific cytosine modifications in the genome without obvious impact on global methylation in 24 h. Finally, we demonstrate our tool can induce targeted CpG methylation in mice by zygote microinjection, thereby demonstrating its potential utility in early development.
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Additional dCas9-tethered domains have been shown to disrupt enhancer activity.
“Additional dCas9-tethered domains have been shown to disrupt enhancer activity (e.g. histone demethylase LSD1210, histone deacetylase 3211, and DNA methylators MQ212 or DNMT3A213-215), and could potentially be adapted to large-scale screens.”
From Towards a comprehensive catalogue of validated and target-linked human enhancers (2020) · cites this paper as [212]
The Methods (Plasmid construction) identify the paper's dCas9–MQ1 fusion, and the Results (Targeted CpG methylation affects CTCF binding) show that its targeted methylation reduced CTCF binding and altered expression at RUNX1. The paper does not show that dCas9–MQ1 disrupted enhancer activity; it tests CTCF-binding sites instead. Page numbers are unavailable in our copy. (LLM, conf=0.83)
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- Authors
- Lei, Y., Zhang, X., Su, J., Jeong, M., Gundry, M. C., Huang, Y.-H., Zhou, Y., Li, W., & Goodell, M. A.
- Affiliations
- Baylor College of Medicine (US)
- Texas A&M University (US)
- Corresponding author
- Goodell, Margaret A.
- ORCID
- Venue
- Nature Communications
- Published
- 2017-07-11
- Volume
- 8
- Issue
- 1
- Pages
- 16026-16026
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC5508226
- OpenAlex
- W2735350095
- Cited by
- 218
- References
- 40
- Citations by year
- 2026: 4 · 2025: 20 · 2024: 22 · 2023: 28 · 2022: 26 · 2021: 26 · 2020: 32 · 2019: 24 · 2018: 32 · 2017: 3
- FWCI
- 10.28
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Nature Communications (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Welch Foundation
- Cancer Prevention and Research Institute of Texas (CA183252, DK092883, BE-1913, CA125123)
- Adrienne Helis Malvin Medical Research Foundation
- Edward P. Evans Foundation
- Evans Medical Foundation
- National Institutes of Health (DK092883, CA183252, DK092883, CA125123, CA193466)
- National Human Genome Research Institute (R01HG007538)
- National Cancer Institute (F99CA222736, K00CA222736, R01CA183252, R01CA193466, P30CA125123, P50CA126752)
- National Institute of General Medical Sciences (R01GM112003)
- National Institute of Diabetes and Digestive and Kidney Diseases (R56DK092883, T32DK060445, R01DK092883)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Keywords
- targeted DNA methylation, cytosine methylation, CpG methylation, zygote microinjection, in vivo gene editing, embryonic development, methylome
- MeSH terms
- Gene Editing, CCCTC-Binding Factor, Animals, Humans, Microinjections, Tenericutes, Recombinant Fusion Proteins, DNA Modification Methylases, CpG Islands, DNA Methylation, K562 Cells, Mice, HEK293 Cells
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