KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade | Abcam | Cat#: ab8898 |
Anti-CBX1 / HP1 beta antibody | Abcam | Cat#: ab10478 |
Anti-KAP1 antibody | Abcam | Cat#: ab10483 |
Alexa Fluor® 647 Donkey Anti-Rabbit IgG (H+L) Antibody | Life Technologies | Cat#: A31573 |
Chemicals, Peptides, and Recombinant Proteins | ||
TransIT-LT1 Transfection Reagent | Mirus | Cat#: MIR 2306 |
Lentivirus Precipitation Solution | Alstem | Cat#: VC100 |
Polybrene | EMD Millipore | Cat#: TR-1003-G |
Abscisic acid | Sigma | Cat#: A1049 |
JF549-HaloTag ligand | Grimm, et al 2015 | N/A |
1,6 hexanediol | Sigma | Cat#: 240117 |
ProLong™ Live Antifade Reagent | ThermoFisher | Cat#: P36975 |
Electroporation enhancer | IDT | Cat#: 1075915 |
Hydroxyurea | Sigma | Cat#: H8627 |
Hoechst 33342 | ThermoFisher | Cat#: H3570 |
Dynabeads Protein A | ThermoFisher | Cat#: 10001D, Lot 00531421 |
Critical Commercial Assays | ||
Qubit dsDNA HS Assay Kit | Invitrogen | Cat #: Q32854 |
SYBR Green Max Mix | BioRad | Cat #: 172-5125 |
Deposited Data | ||
H3K9me3 ChIP-seq | GEO | GSE175500 |
Experimental Models: Cell Lines | ||
Human: U-2 OS cell line | ATCC | Cat#: HTB-96 |
Human: LentiX 293T | Takara | Cat#: 632180 |
Oligonucleotides | ||
sgRNAs for Cas9 | See Table S2 for sequences and information. | N/A |
Fluorescently labeled crRNAs for Cas9 | See Table S3 for sequences and information. | N/A |
RT-qPCR primers | See Table S4 for sequences and information. | N/A |
Recombinant DNA | ||
pHR: pmU6 sgRNA pCMV Puro | This paper | pSLQ1844 |
pHR: pTRE3G dCas9-2xNLS-HaloTag | Wang et. al. 2018 | pSLQ4218 |
pHR: pPGK ABI-tagBFP-dCas9-2xNLS | Wang et. al. 2018 | pSLQ4356 |
pHR: pPGK PYL1-sfGFP-HP1A | Wang et. al. 2018 | pSLQ6901 |
pHR: pPGK PYL1-sfGFP-HP1Adel72 | This paper | pSLQ6903 |
pHR: pPGK PYL1-sfGFP-HP1AI165E | This paper | pSLQ6904 |
pHR: pPGK mCherry-HP1A | This paper | pSLQ6909 |
pHR: pPGK PYL1-sfGFP-NLS | This paper | pSLQ6918 |
pHR: pmU6 sgRNA 2xMS2 pUbC MCP-mCherry-p2a-Puro | This paper | pSLQ6941 |
Software and Algorithms | ||
Fiji (ImageJ) | Schindelin et al. 2012 | https://fiji.sc/ |
TrackMate plugin | Tinevez et. al. 2017 | https://imagej.net/TrackMate |
Correct 3D Drift plugin | Parslow et. al. 2014 | https://imagej.net/Correct_3D_Drift |
Bleach correction macro | J. Rietdorf, EMBL Heidelberg | https://imagej.net/Bleach_Correction |
NIS Elements Version 4.60 | Nikon | https://www.microscope.healthcare.nikon.com/products/software/nis-elements |
Prism 8 | GraphPad | https://www.graphpad.com/ |
FlowJo | FlowJo, LLC | https://www.flowjo.com/ |
Bowtie2 | Langmead et. al. 2012 | http://bowtie-bio.sourceforge.net/bowtie2/index.shtml |
Samtools | Whitman Institute | http://www.htslib.org/ |
Interactive Genome Viewer | Broad Institute | https://software.broadinstitute.org/software/igv/ |
bigwiggeneration.txt | Zenodo | https://doi.org/10.5281/zenodo.5110819 |
Other | ||
QSonica Q500 | QSonica | https://www.sonicator.com/ |