Journal: Cell

Article Title: Rescue of Fragile X syndrome neurons by DNA methylation editing of the FMR1 gene

doi: 10.1016/j.cell.2018.01.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Lentiviruses labeling NPCs (EF1A-GFP and EF1A-RFP) were purchased from Cellomics Technology. table ft1 table-wrap mode="anchored" t5 caption a7 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-Cas9 (IF staining) Active Motif Cat#61577 (7A9-3A3) Mouse monoclonal anti-Cas9 (ChIP) Active Motif Cat#61757 (8C1-F10) Chicken polyclonal anti-GFP Aves Labs Cat#GFP-1020 Chicken polyclonal anti-MAP2 Encor Biotech Cat# CPCA-MAP2 Rabbit polyclonal anti-FMRP (WB, ICC, IHC) Cell Signaling Cat#4317 Mouse monoclonal anti-FMRP (ICC, IHC) Biolegend Cat#834701 Mouse monoclonal anti-FMR1PolyG EMD Millipore Cat#MABN784 Goat polyclonal anti-mCherry SICGEN Cat#AB0040-200 Mouse monoclonal anti-RNA Polymerase II Abcam Cat#ab817 Rabbit polyclonal anti-H3K27Ac Abcam Cat#ab4729 Rabbit polyclonal anti-H3K9me3 Abcam Cat#ab8898 Mouse monoclonal anti-H3K27me3 Abcam Cat#ab6002 Rabbit polyclonal anti-H3K4me3 Millipore Cat#07-473 Chemicals Doxycycline hyclate Sigma Aldrich Cat#D9891-100G Critical Commercial Assays EpiTect Bisulfite Kit Qiagen Cat#59104 EpiNext™ High-Sensitivity Bisulfite-Seq kit EPIGENTEK Cat#P-1056 EpiNext™ NGS Barcode Set-12 EPIGENTEK Cat#P-1060 DNeasy Blood & Tissue Kit Qiagen Cat#69504 Zymoclean Gel DNA Recovery Kit Zymo Research Cat#D4002 DNA Clean & Concentrator-5 Zymo Research Cat#D4013 X-tremeGENE™ 9 DNA Transfection Reagent Sigma Aldrich Cat#6365809001 Direct-zol RNA Miniprep Zymo Research Cat#R2050 SuperScript III First-Strand Synthesis SuperMix Life Technologies Cat#18080400 Fast SYBR Green Master Mix Life Technologies Cat#4385618 Deposited Data Raw data files for sequencing NCBI Gene Expression Omnibus Database: {"type":"entrez-geo","attrs":{"text":"GSE102655","term_id":"102655"}} GSE102655 , {"type":"entrez-geo","attrs":{"text":"GSE102684","term_id":"102684"}} GSE102684 , {"type":"entrez-geo","attrs":{"text":"GSE108171","term_id":"108171"}} GSE108171 , {"type":"entrez-geo","attrs":{"text":"GSE108498","term_id":"108498"}} GSE108498 , {"type":"entrez-geo","attrs":{"text":"GSE108577","term_id":"108577"}} GSE108577 Experimental Models: Organisms/Strains Mouse: NOD- scid IL2Rg null , Jackson Laboratories Stock No: 005557 Experimental Models: Cell Lines FX52 iPSCs ( Urbach et al., 2010 ) N/A FXS iPSC (135.3) ( Brick et al., 2014 ) N/A FXS iPSC (FXS_SW) ( Xie et al., 2016 ) N/A Recombinant DNA Fuw-dCas9-Tet1-P2A-BFP This paper N/A pgRNA This paper N/A Fuw-AcrIIA4-P2A-GFP This paper N/A See “ Plasmid design and construction ” in METHOD DETAILS section Sequence-Based Reagents See Table S4 , S5 , S6 for primer sequences This paper N/A Software and Algorithms MACS (ChIP-seq algorithms) Wu et al., 2014b http://liulab.dfci.harvard.edu/MACS/ ImageJ (Fiji) NIH http://imagej.net/Fiji Open in a separate window

Techniques: Staining, Bisulfite Sequencing, Transfection, SYBR Green Assay, Sequencing, Expressing, Recombinant, Plasmid Preparation, Software