DYS‐HAC2 transfer into reversibly immortalised DMD myoblasts

Slides:

Advertisements

Similar presentations

Rupp et al. Supplementary Figure 1: Structure of the human troponin T gene Exon 6 Genomic DNA cDNA from mRNA mutation Exon 9 Exon bp Parts of genomic.

Advertisements

Volume 17, Issue 2, Pages (August 2015)

Statistical analysis of transgene expression in pigs generated by lentiviral gene transfer. Statistical analysis of transgene expression in pigs generated.

Loss of Hdac3 impairs Ar signaling in mouse prostate tissues and has no effect on apoptosis Loss of Hdac3 impairs Ar signaling in mouse prostate tissues.

DNA‐anchored LacI–CFP–Luciferase aggregates capture misfolded cytosolic proteins. DNA‐anchored LacI–CFP–Luciferase aggregates capture misfolded cytosolic.

REF and ATP5b status on mitochondrial fraction.

Increased proliferation and delayed differentiation of adult primary myoblasts derived from MyoDR99,102 mice. Increased proliferation and delayed differentiation.

Vav‐1 gene‐targeting strategy.

GLUT3 is a direct target of TCF4/β‐catenin

c‐Abl acetylation on Lys 730 promotes myogenic differentiation.

Isolation of genetically corrected recessive dystrophic epidermolysis bullosa (RDEB) epidermal stem cellsSingle cells were isolated from a mass culture.

AAV8‐hAAT‐FGF21 treatment improves liver fibrosis

Dual LGR5 and KI67 knock‐in PDOs enable separation of quiescent and cycling LGR5+ CRC cells Dual LGR5 and KI67 knock‐in PDOs enable separation of quiescent.

Characterisation of proviral integrations in corrected stem cells Visualisation of COL7A1 sequence by FISH analysis in corrected stem cells at passage.

VAP‐A is recruited to interorganelle contact sites together with MOSPD2 VAP‐A is recruited to interorganelle contact sites together with MOSPD2 AHeLa cells.

Generation of a novel, synthetic, multifunctional DYS‐HAC

TGF‐β1 and Sema3A are downregulated in vivo by increasing VEGF doses Muscles were harvested 7 days after implantation of V Low, V Med, and V High clones.

Lineage tracing of LGR5+ CRCs in human colorectal xenografts

Volume 18, Issue 4, Pages (April 2016)

Modules with causal links to ALS are associated with inflammation and enriched with TDP‐43 protein–protein interactions (PPIs)‏ Modules with causal links.

HPV‐E7/RB axis regulates ceramide‐dependent mitophagy via E2F5 activation HPV‐E7/RB axis regulates ceramide‐dependent mitophagy via E2F5 activation AUM‐SCC‐47.

KLK7 is involved in the Aβ degradation activity

Expression of PD‐1 on PBMC and malignant exudate T cells

Expression and occupancy of a set of transcription factors corresponding to the identified motifs at FAIRE peaks Expression and occupancy of a set of transcription.

Integrated protein co‐expression and differential expression across the ALS‐FTD disease continuum Integrated protein co‐expression and differential expression.

MOSPD2 is a putative tail‐anchored protein

Isolation of genetically corrected recessive dystrophic epidermolysis bullosa (RDEB) epidermal stem cellsSingle cells were isolated from a mass culture.

Superior potency of EnAd expressing EpCAM BiTE in partially EnAd‐resistant cancer cell line Superior potency of EnAd expressing EpCAM BiTE in partially.

Expression of GFP‐Aub transgenes in GSCs, and GSC loss phenotype in armi mutant Expression of GFP‐Aub transgenes in GSCs, and GSC loss phenotype in armi.

Fig. 1. Generation of the ΔEx50 mouse model.

BRD4 expression levels are associated with sensitivity to BET inhibition in NRAS‐mutant melanoma BRD4 expression levels are associated with sensitivity.

Validation of astrocyte and microglial proteomic markers that increase across the ALS‐FTD spectrum Validation of astrocyte and microglial proteomic markers.

Volume 17, Issue 2, Pages (August 2015)

FFAT‐containing proteins recruit the ER‐resident MOSPD2 protein to interorganelle contact sites FFAT‐containing proteins recruit the ER‐resident MOSPD2.

Reduced ALPI expression in patients’ small intestinal biopsies

Detection and tracking of individual SNAP-tagged proteins on the surface of living cells. Detection and tracking of individual SNAP-tagged proteins on.

Nec‐1 inhibits the phosphorylation and aggregation of tau

Hdac3 deletion decreases AKT phosphorylation and tumor growth in Pten knockout prostate cancer Hdac3 deletion decreases AKT phosphorylation and tumor growth.

NILV‐S/MAR‐mediated transgene expression

TGF‐β1 treatment induces the chromatin binding of Smad2/3/4 in 4T1 cells TGF‐β1 treatment induces the chromatin binding of Smad2/3/4 in 4T1 cells A, BWestern.

BDNF expression in the cerebellum and brain stem region.

Boxplots representing the seven protein profiles significantly differing between muscular dystrophy patients and control groupsEach boxplot represents.

Cochlear explant and slice preparation, and CDDP‐induced DNA damage and cell death in sensory hair cells and spiral ganglion neurons Cochlear explant and.

Safety profile of reversibly immortalised human mesoangioblasts

EphB4 stimulation does not cause vessel regression

EnAd‐EpCAMBiTE shows reproducible activity within an expanded cohort of patient malignant peritoneal and pleural exudates EnAd‐EpCAMBiTE shows reproducible.

Cbl protein levels in ago3, twin, and armi mutant GSCs

Distinct collagen structures in the upper and lower neonatal dermis (related to Fig 1)‏ Distinct collagen structures in the upper and lower neonatal dermis.

Validation of astrocyte and microglial proteomic markers that increase across the ALS‐FTD spectrum Validation of astrocyte and microglial proteomic markers.

Assessment of proliferation status of PDGFRα+ cells in developing mouse lungs Assessment of proliferation status of PDGFRα+ cells in developing mouse lungs.

Assessment of immortalisation, tumorigenicity and disseminative potential of the corrected stem cells Upper panel: expression of proteins associated with.

ZBTB48 is required for MTFP1 expression

Drp1 knockdown prevents mitophagy

Microglial P2X4R modulates oligodendrocyte differentiation

Re‐expression of the AXIN1 coding sequence re‐sensitizes VACO6R cells to PORCN inhibition Re‐expression of the AXIN1 coding sequence re‐sensitizes VACO6R.

ALK5 inhibition induces ubiquitin‐mediated degradation of Smad4 in CD8+ T cells in melanoma‐bearing mice Source data is available for this figure in the.

LXRα enhances glucose uptake and O‐GlcNAc signaling via activation of the hexosamine biosynthetic pathway (HBP) in cultured cardiomyocytes LXRα enhances.

T22‐GFP‐H6‐FdU‐induced depletion of CXCR4‐overexpressing cancer cells in tumor tissue T22‐GFP‐H6‐FdU‐induced depletion of CXCR4‐overexpressing cancer cells.

CRISPR/Cas9-Mediated Deletion of CTG Expansions Recovers Normal Phenotype in Myogenic Cells Derived from Myotonic Dystrophy 1 Patients Claudia Provenzano,

GSC tumor phenotype in ago3 mutant germaria

DIA1(R1204X) induces elongated and thick microvilli in HeLa cells

Effect of D‐Asp treatment on myelination and remyelination in cerebellar organotypic slices Effect of D‐Asp treatment on myelination and remyelination.

NILV‐S/MAR‐mediated transgene expression

GDF11 induces myotube atrophy in vitro

Tangle formation and downregulation of miR‐132‐3p in LOAD A‐G

Dnmt2 mutants show delayed immune responses and Dnmt2–EGFP relocalizes and interacts with DCV RNA during infection. Dnmt2 mutants show delayed immune responses.

UCN‐01 inhibits human IPF lung fibroblast pathological phenotypes and attenuates lung fibrosis induced by bleomycin injury UCN‐01 inhibits human IPF lung.

Fig. 1 Exon 44–deleted DMD patient iPSC-derived cardiomyocytes express dystrophin after CRISPR-Cas9–mediated genome editing. Exon 44–deleted DMD patient.

Fig. 2 DMD iPSC-derived cardiomyocytes express dystrophin after Cpf1-mediated genome editing by reframing. DMD iPSC-derived cardiomyocytes express dystrophin.

Fig. 3 Correction of Dmd exon 44 deletion in mice by intramuscular AAV9 delivery of gene editing components. Correction of Dmd exon 44 deletion in mice.

Presentation transcript:

DYS‐HAC2 transfer into reversibly immortalised DMD myoblasts DYS‐HAC2 transfer into reversibly immortalised DMD myoblasts PCR panel for HAC sequences in four neomycin‐resistant riDMD(DYS‐HAC2) myoblast clones. Red boxes: positive riDMD(DYS‐HAC2) myoblasts.PCR analysis of human dystrophin exons on genomic DNA of riDMD myoblasts, riDMD(DYS‐HAC2)#α and riDMD(DYS‐HAC2)#δ clones. Parental riDMD myoblasts: negative control for exons 5–7; CHO(DYS‐HAC2)‐7: positive control.Left panel: karyotype analysis of riDMD myoblasts, riDMD(DYS‐HAC2)#α and riDMD(DYS‐HAC2)#δ myoblast clones. Right panel: FISH analysis on riDMD myoblasts and two selected riDMD(DYS‐HAC2) myoblast clones (#α and #δ). Insets: magnifications showing single episomal DYS‐HAC2. Red/purple: rhodamine‐p11‐4 human alpha satellite (centromeres of chromosome 13 and 21, hChr 13/21(cen)); green: dystrophin FITC‐DMD‐BAC RP11‐954B16; yellow: merge. White arrowheads: DYS‐HAC2. Scale bar: 3 μm.RT–PCR panel for dystrophin expression in differentiated riDMD(DYS‐HAC2)#α. Parental riDMD myoblasts: negative control for dystrophin (exons 5–7); healthy myoblasts: positive control.Western blot for dystrophin (427 kDa) in differentiated riDMD and riDMD(DYS‐HAC2)#α myoblasts; human muscle and differentiated human inducible myogenic cells (Maffioletti et al, 2015) were used as positive control; differentiated DMD‐inducible myogenic cells (Maffioletti et al, 2015) were used as negative control and myosin heavy chain (MyHC) as normaliser (40 μg of proteins loaded for all samples but human muscle, which had 30 μg loaded).Immunofluorescence images showing in vitro muscle differentiation of riDMD myoblasts (negative control), riDMD(DYS‐HAC2)#α and healthy donor myoblasts (positive control). Red: MyHC; green: dystrophin; blue: Hoechst. Scale bar: 50 μm. Source data are available online for this figure. Sara Benedetti et al. EMBO Mol Med. 2017;emmm.201607284 © as stated in the article, figure or figure legend