jds246 addgene  (Addgene inc)

Journal: Cell

Article Title: A high-throughput platform to identify small-molecule inhibitors of CRISPR-Cas9

doi: 10.1016/j.cell.2019.04.009

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Table S1 Recombinant DNA pMJ806 Jinek et al ., 2012 Addgene Cat# 39312 JDS246 Fu et al ., 2013 Addgene Cat# 43861 pFYF1320 EGFP SiteCat# 1 Fu et al ., 2013 Addgene Cat# 47511 BPK1520 Kleinstiver et al ., 2015 Addgene Cat# 65777 MS2-P65-HSF1 Maji et al ., 2017 NA dSpCas9 Feng Zhang lab NA Critical Commercial Assays MEGAClear Transcription Clean Up Kit RNA purification Thermo Fisher Cat# AM1908 High Precision Streptavidin (SAX) Biosensors ForteBio Cat# 18–5117 SE Cell Line 4D-Nucleofector X Kit Lonza Cat# V4XC-1032 Qubit dsDNA HS Kit Thermo Fisher Cat# {"type":"entrez-protein","attrs":{"text":"Q32851","term_id":"75280859","term_text":"Q32851"}} Q32851 Nano-Glo HiBiT Lytic Reagent Promega Cat# N3030 TaqMan probes Life Technologies Cat# 4331182 QuickExtract Epicentre Cat# QE09050 Surveyor Mutation Detection Kits IDT Cat# 706020 MiSeq Reagent Kit v2 300 Illumina Cat# MS-102–2002 Streptavidin magnetic beads Thermo Fisher Scientific Cat# 88816 RevertAid RT Reverse Transcription Kit Thermo Fischer Scientific Cat# K1691 Ni-NTA agarose resin Thermo Fisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"R90110","term_id":"957650","term_text":"R90110"}} R90110 QIAamp DNA Mini Kit Qiagen Cat# 51306 Software and Algorithms GraphPad Prism GraphPad Prism Software Inc. https://www.graphpad.com/scientific-software/prism/ Spotfire Tibco https://www.tibco.com/products/tibco-spotfire MetaXpress Molecular Device https://www.moleculardevices.com/products/cellular-imaging-systems/acquisition-and-analysis-software/metaxpress#gref MATLAB MathWorks https://in.mathworks.com/ MATLAB Script Komor et al ., 2016 N/A Open in a separate window KEY RESOURCES TABLE Developed high-throughput assays for SpCas9 and performed a small-molecule screen Identified reversible and cell-permeable inhibitors that disrupt SpCas9-DNA binding Inhibitors allow dose and temporal control of (non)-nuclease-based SpCas9 systems Identified a pharmacophore for SpCas9 inhibition using structure-activity studies

Techniques: Recombinant, Cell Culture, Western Blot, Labeling, Purification, Mutagenesis, Magnetic Beads, Software