Evidence map›Paper›PMID 28751638›Full record

ArticleScientific reports2017

Targeted DNA methylation in human cells using engineered dCas9-methyltransferases.

Tina Xiong, Glenna E Meister, Rachael E Workman, Nathaniel C Kato, Michael J Spellberg, Fulya Turker, Winston Timp, Marc Ostermeier, Carl D Novina

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 53 papers.

0numbers the graph read from it
0cells of the map it votes in
53citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

53 citing papers in PubMed.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

9 authors.

Tina XiongDepartment of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
Glenna E MeisterDepartment of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.
Rachael E WorkmanDepartment of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
Nathaniel C KatoDepartment of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
Michael J SpellbergDepartment of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.
Fulya TurkerDepartment of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.
Winston TimpDepartment of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
Marc OstermeierDepartment of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA. oster@jhu.edu.
Carl D NovinaDepartment of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA. carl_novina@dfci.harvard.edu.

Funding

Engineering epigenetic therapy for sickle cell diseaseDP1DK105602 · NIDDK · DANA-FARBER CANCER INST · PI NOVINA, CARL D · 2014 to 2018
$4.3M
NIDDK NIH HHS DP1 DK105602
6 · The paper itself

Abstract

Mammalian genomes exhibit complex patterns of gene expression regulated, in part, by DNA methylation. The advent of engineered DNA methyltransferases (MTases) to target DNA methylation to specific sites in the genome will accelerate many areas of biological research. However, targeted MTases require clear design rules to direct site-specific DNA methylation and minimize the unintended effects of off-target DNA methylation. Here we report a targeted MTase composed of an artificially split CpG MTase (sMTase) with one fragment fused to a catalytically-inactive Cas9 (dCas9) that directs the functional assembly of sMTase fragments at the targeted CpG site. We precisely map RNA-programmed DNA methylation to targeted CpG sites as a function of distance and orientation from the protospacer adjacent motif (PAM). Expression of the dCas9-sMTase in mammalian cells led to predictable and efficient (up to ~70%) DNA methylation at targeted sites. Multiplexing sgRNAs enabled targeting methylation to multiple sites in a single promoter and to multiple sites in multiple promoters. This programmable de novo MTase tool might be used for studying mechanisms of initiation, spreading and inheritance of DNA methylation, and for therapeutic gene silencing.

Indexed as

DNA MethylationBase SequenceBinding SitesCpG IslandsCRISPR-Associated Protein 9DNA (Cytosine-5-)-MethyltransferasesEscherichia coliGene EditingGenetic VectorsHEK293 CellsHumansKineticsModels, MolecularProtein BindingProtein Conformation, alpha-HelicalProtein Conformation, beta-StrandCas9 endonuclease Streptococcus pyogenesCRISPR-Associated Protein 9DNA (Cytosine-5-)-MethyltransferasesRecombinant Fusion ProteinsRNA, Guide, CRISPR-Cas Systems

Identifiers

PMID28751638
PMCPMC5532369

What OpenQuestion holds

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LicenceCC BY
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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.