KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
| ||
Anti-Histone H3 (acetyl K27) - ChIP Grade | Abcam | ab4729, Lot: GR3374555-1 |
Anti-Histone H3 (mono methyl K4) | Abcam | ab8895, Lot: GR3407156-1 |
Anti-OTX2 (D7Y3J) | Cell Signaling Technology | mAb #11943, Lot#1 |
Anti-Nanog (A-11) | Santa Cruz Biotechnology | sc-374001, Lot#E0521 |
Anti-PAX7 | DSHB | N/A |
β-Actin (N-21) | Santa Cruz Biotechnology | sc-130656, Lot #C2515 |
MyoD (4H207) | Santa Cruz Biotechnology | sc-71629, Lot #G2012 |
MyoG (F5D) | Santa Cruz Biotechnology | sc-12732, Lot #A0620 |
Goat Anti Rabbit IgG-HRP | Azure biosystem | AC2114, Lot 210812-57 |
Goat Anti mouse IgG-HRP | Azure biosystem | AC2115, Lot 210204-50 |
| ||
Bacterial and virus strains | ||
| ||
NEB® 5-alpha Competent E. coli (High Efficiency) | NEB | C2987H |
One Shot™ TOP10 Chemically Competent E. coli | ThermoFisher scientific | C404010 |
| ||
Chemicals, peptides, and recombinant proteins | ||
| ||
Knock-Out™ Serum Replacement | ThermoFisher | 10828028 |
Recombinant Protein-Rspo3 | R&D Biosystems | 4120-RS-025 |
Leukemia inhibitory factor-LIF | Millipore | ESG1106 |
Recombinant Murine BMP4 | Peprotech | 315–27 |
BSA solution | Gibco | 101020–021 |
LDN193189 | Stemgent | 04–0074 |
CHIRON99021 | Stemgent | 04-0004-02 |
Recombinant Murine FGF | Peprotech | 450-33 |
Recombinant Murine IGF | Peprotech | 250-19 |
Recombinant Mouse HGF | R&D Biosystems | 2207-HG-2-025 |
Dynabeads Protein G | ThermoFisher Scientific | 10004D |
TRIzol Reagent | Thermo Fisher Scientific | 15596018 |
Polybrene | Sigma | TR-1003-G |
Protease Inhibitor | Roche | 11836170001 |
Phase Lock Gel Heavy | 5 PRIME | 2302810 |
Chloroform | Sigma | 67-66-3 |
10% SDS | Invitrogen | 15553-027 |
Neurobasal Medium | Gibco | 21103-049 |
DMEM/F12 (1:1) | Gibco | 11320-033 |
Dimethyl sulfoxide | Sigma | 276855 |
2-Mercaptoethanol | Sigma | M6250 |
Dulbecco’s Modified Eagle Medium (DMEM) | Gibco | 12439-054 |
Pen Strep Glutamine (100x) | Gibco | 10378-016 |
GlutaMAX (100x) | Gibco | 35050-061 |
MEM NEAA | Gibco | 11140-050 |
Sodium Pyruvate (100mM) | Gibco | 11360-070 |
BSA Fraction V (7.5%) | Gibco | 15260-037 |
| ||
Critical commercial assays | ||
| ||
DNeasy Blood & Tissue Kits | Qiagen | 69504 |
QIAprep Spin Miniprep Kit | Qiagen | 27106 |
Lenti-X™ Concentrator | Clontech | 631231 |
Lipofectamine 2000 | Invitrogen | 11668-019 |
Power SYBR Green PCR Master Mix | Thermo Fisher Scientific | 4368708 |
PCR purification kit | Qiagen | 28106 |
qScript™ cDNA Synthesis Kit | Quantabio | 95047-100 |
Arima HiC kit | Arima | A510008GFP |
Accel-NGSO 2S Plus DNA | SWIFT Bioscience | A160140 v00 |
NEBNext Ultra II DNA library prep kit | NEB | #E7645 |
NEBNext Ultra II RNA library preparation kit for Illumina | NEB | #E7490 |
Chromium Single Cell 3’ Library & Gel Bead Kit v3.1 | 10x Genomics | P/N 1000121 |
| ||
Deposited data | ||
| ||
RNA-seq, ATAC-seq, H327ac and H3K4me1 ChIP-seq, HiC, Single cell RNA-seq, Single cell ATAC-seq, Multiome single nuclei ATAC and gene expression | This paper | GSE198730 |
| ||
Experimental models: Cell lines | ||
| ||
HEK 293T cells | ATCC | CRL-1573 |
C2C12 cells | ATCC | GSC-6001G |
ESC Pax3-GFP | Chal et al., 2015 | N/A |
ESC Pax3-GFP Del −3.5kb | This paper | N/A |
ESC Pax3-GFP Del −3.5kb and −25kb | This paper | N/A |
ESC Pax3-GFP Del En7 | This paper | N/A |
| ||
Oligonucleotides | ||
| ||
sgRNA sequences | Table S4 | N/A |
RT q-PCR oligos | Table S4 | N/A |
PCR oligos | Table S4 | N/A |
| ||
Recombinant DNA | ||
| ||
pSpCas9(BB)-2A-GFP (PX458) | Addgene | Plasmid #48138 |
pSpCas9(BB)-2A-GFP-gRNA1_−3.5kb | This paper | N/A |
pSpCas9(BB)-2A-GFP-gRNA4_−3.5kb | This paper | N/A |
pSpCas9(BB)-2A-GFP-gRNA1_−25kb | This paper | N/A |
pSpCas9(BB)-2A-GFP-gRNA4_−25 kb | This paper | N/A |
pSpCas9(BB)-2A-GFP-gRNA2_En7 | This paper | N/A |
pSpCas9(BB)-2A-GFP-gRNA4_En7 | This paper | N/A |
pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-GFP | Addgene | Plasmid #71237 |
pLV hU6-sgRNA_Pax7_promoter_ hUbC-dCas9-KRAB-T2a-GFP | This paper | N/A |
pLV hU6-sgRNA_Pax7_−3.5kb_ hUbC-dCas9-KRAB-T2a-GFP | This paper | N/A |
pLV hU6-sgRNA_Pax7_−25kb_ hUbC-dCas9-KRAB-T2a-GFP | This paper | N/A |
pLV hU6-sgRNA_Pax7_ En7_ hUbC-dCas9-KRAB-T2a-GFP | This paper | N/A |
psPAX2 | Addgene | Plasmid ##12260 |
pMD2.G | Addgene | Plasmid #12259 |
pGL4.26[luc2/minP/Hygro] | Promega | #E8441 |
pGL4.26-Ctrl | This paper | N/A |
pGL4.26-En7 | This paper | N/A |
| ||
Software and algorithms | ||
| ||
BioRender | biorender.com | |
Prism Software, version 8 | Graphpad Software | https://www.Graphpad.com |
Metascape | Zhou et al. (2019) | https://metascape.org/gp/index.html#/main/step1 |
CellRanger ver. 3.1.0 | 10x Genomics | https://support.10xgenomics.com/single-cell-gene-expression/software/pipelines/latest/installation |
Cell Ranger ATAC v1.1.0 | 10x Genomics | https://support.10xgenomics.com/single-cell-atac/software/pipelines/latest/what-is-cell-ranger-atac |
R | The R Project for Statistical Computing | https://www.r-project.org/ |
Seurat v4.1.0 | Stuart et al., 2019 | https://cran.r-project.org/web/packages/Seurat/index.html |
Signac v1.5.0 | Stuart et al., 2021 | https://satijalab.org/signac |
Harmony | Korsunsky et al., 2019 | https://github.com/immunogenomics/harmony |
Monocle3 | Cao et al., 2019 | https://cole-trapnell-lab.github.io/monocle3/ |
JASPAR 2020 | Fornes et al. (2020) | https://bioconductor.org/packages/release/data/annotation/html/JASPAR2020.html |
FBSTools | Tan and Lenhard (2016) | https://bioconductor.org/packages/release/bioc/html/TFBSTools.html |
cellranger-arc version 2.0.0 | 10x Genomics | https://support.10xgenomics.com/single-cell-multiome-atac-gex/software/pipelines/latest/what-is-cell-ranger-arc |
Seurat ver. 4.1.0 | Hao et al., 2021 | https://satijalab.org/seurat/ |
Signac ver.1.5.0 | Stuart et al., 2021 | https://satijalab.org/signac/news/index.html |
MACS 1.4.2 | Zhang et al., 2008 | https://libraries.io/pypi/MACS |
Bowtie/1.1.1 | Langmead et al., 2009 | http://bowtie-bio.sourceforge.net/index.shtml |
BedGraphToBigWig | Kent et al., 2010 | https://www.encodeproject.org/software/bedgraphtobigwig/ |
Bedtools/2.29.2 | Quinlan and Hall, 2010 | https://bedtools.readthedocs.io/en/latest/content/installation.html |
HOMER/4.11.1 | Heinz et al., 2010 | http://homer.ucsd.edu/homer/ |
Partek Genomics Suite 7.18 | Partek Inc. | https://www.partek.com/partek-genomics-suite/ |
TopHat 2.1.1 | Trapnell et al., 2009 | https://ccb.jhu.edu/software/tophat/index.shtml |
Trim Galore (version 0.6.6) | N/A | https://github.com/FelixKrueger/TrimGalore |
Bcl2fastq/2.20.0 | Illumina | https://emea.support.illumina.com/downloads/bcl2fastq-conversion-software-v2-20.html |
Juicer 1.6 | Durand et al., 2016 | https://github.com/aidenlab/juicer/releases |
HiCExplorer3.6 | Wolff et al., 2020 | https://hicexplorer.readthedocs.io/en/latest/ |
HOMER | Heinz et al., 2010 | http://homer.ucsd.edu/homer/ |
Custom scripts | This paper | https://doi.org/10.5281/zenodo.6889278 |