Generation of a transgene-free iPSC line and genetically modified line from a facioscapulohumeral muscular dystrophy type 2 (FSHD2) patient with SMCHD1 p.Lys607Ter mutation

Sasaki-Honda, Mitsuru; Kagita, Akihiro; Jonouchi, Tatsuya; Araki, Toshiyuki; Hotta, Akitsu; Sakurai, Hiroyuki

doi:10.1016/j.scr.2020.101884

Cited by 2 publications

(2 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…iPS cell clones with Tetracycline (TET)-inducible myogenic potential were previously established from a patient with FSHD1, a patient with FSHD2 and a healthy control donor ( 43 ). Myocyte differentiation was conducted as previously described with minor modifications ( 43 , 44 ). Duration and concentration of doxycycline stimulation was optimized for each iPS cell clone, from 4 to 5 days and from 0.3 to 1.0 μg/ml, respectively, to achieve comparable myogenic differentiation.…”

Section: Methodsmentioning

confidence: 99%

Nanopore direct RNA sequencing detects DUX4-activated repeats and isoforms in human muscle cells

Mitsuhashi

Nakagawa

Sasaki-Honda

et al. 2021

Human Molecular Genetics

Self Cite

View full text Add to dashboard Cite

Facioscapulohumeral muscular dystrophy (FSHD) is an inherited muscle disease caused by misexpression of the DUX4 gene in skeletal muscle. DUX4 is a transcription factor which is normally expressed in the cleavage-stage embryo and regulates gene expression involved in early embryonic development. Recent studies revealed that DUX4 also activates the transcription of repetitive elements such as endogenous retroviruses (ERVs), mammalian apparent LTR-retrotransposons (MaLRs), and pericentromeric satellite repeats (HSATII). DUX4-bound ERV sequences also create alternative promoters for genes or long non-coding RNAs (lncRNAs), producing fusion transcripts. To further understand transcriptional regulation by DUX4, we performed nanopore long-read direct RNA sequencing (dRNA-seq) of human muscle cells induced by DUX4, because long reads show whole isoforms with greater confidence. We successfully detected differential expression of known DUX4-induced genes, and discovered 61 differentially-expressed repeat loci, which are near DUX4-ChIP peaks. We also identified 247 gene-ERV fusion transcripts, of which 216 were not reported previously. In addition, long-read dRNA-seq clearly shows that RNA splicing is a common event in DUX4-activated ERV transcripts. Long-read analysis showed non-LTR transposons including Alu elements are also transcribed from LTRs. Our findings revealed further complexity of DUX4-induced ERV transcripts. This catalogue of DUX4-activated repetitive elements may provide useful information to elucidate the pathology of FSHD. Also, our results indicate that nanopore dRNA-seq has complementary strengths to conventional short read cDNA sequencing.

show abstract

Section: Methodsmentioning

confidence: 99%

Nanopore direct RNA sequencing detects DUX4-activated repeats and isoforms in human muscle cells

Mitsuhashi

Nakagawa

Sasaki-Honda

et al. 2021

Human Molecular Genetics

Self Cite

View full text Add to dashboard Cite

show abstract

“…[83] The authors further published the generation of a transgene-free iPSC line and genetically modified line from a FSHD2 patient with the p.Lys607Ter mutation in SMCHD1. [84] The current consensus for FSHD pathogenesis is that the loss of epigenetic repression of the D4Z4 macrosatellite repeat (and hence of the DUX4 locus) prominently, if not fundamentally, underlies the pathogenesis of both FSHD1 and FSHD2. Many studies have recently contributed towards the understanding of the genes regulating DUX4 expression [37,39,55] and the downstream consequences of DUX4 activation.…”

Section: Insights Into the Neurobiology Of Fshd2mentioning

confidence: 99%

Facioscapulohumeral muscular dystrophy type 2: an update on the clinical, genetic, and molecular findings

Jia

Drew

Nicholson

et al. 2021

Neuromuscular Disorders

View full text Add to dashboard Cite

This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

show abstract

Generation of a transgene-free iPSC line and genetically modified line from a facioscapulohumeral muscular dystrophy type 2 (FSHD2) patient with SMCHD1 p.Lys607Ter mutation

Cited by 2 publications

References 4 publications

Nanopore direct RNA sequencing detects DUX4-activated repeats and isoforms in human muscle cells

Nanopore direct RNA sequencing detects DUX4-activated repeats and isoforms in human muscle cells

Facioscapulohumeral muscular dystrophy type 2: an update on the clinical, genetic, and molecular findings

Contact Info

Product

Resources

About