Antibodies |
Mouse monoclonal anti-β-Actin, HRP-linked |
Sigma-Aldrich |
Cat# A3854-200UL |
Mouse monoclonal anti-FLAG |
Sigma-Aldrich |
Cat# F1804 |
Rabbit polyclonal anti-H3K4me3 |
Sigma-Aldrich |
Cat# 07–473 |
Rat monoclonal anti-HA (clone 3F10), HRP-linked |
Roche |
Cat# 12013819001 |
Rabbit monoclonal anti-SCP4 |
Cell Signaling Technology |
Cat# 6932 |
Rabbit polyclonal anti-mouse IgG, HRP-linked |
Agilent/Dako |
Cat# P026002-2 |
Donkey anti-rabbit IgG, HRP-linked |
Cytiva/Amersham |
Cat# NA934 |
Bacterial and virus strains |
MegaX DH10B T1R Electrocomp Cells |
Invitrogen |
Cat# C640003 |
Chemicals, peptides, and recombinant proteins |
2-Mercaptoethanol |
Sigma-Aldrich |
Cat# M6250 |
2x Laemmli Sample Buffer |
BIO-RAD |
Cat# 1610737 |
Agarose, Standard, Low Electroendosmosis (EEO) |
Avantor |
Cat# A426-07 |
Anti-FLAG ® M2 Affinity Gel |
Sigma-Aldrich |
Cat# A2220-10ML |
Blasticidin S HCl |
Thermo Fisher Scientific |
Cat# A1113903 |
Blasticidin S HCl |
Thermo Fisher Scientific |
Cat# A1113903 |
BsmBI |
New England BioLabs |
Cat# R0739 |
Cell Lysis Buffer |
Cell Signaling Technology |
Cat# 9803 |
Chloroform |
Sigma-Aldrich |
Cat# 288306 |
D-Luciferin |
Goldbio |
Cat# 115144-35-9 |
Dimethyl Sulfoxide |
Sigma-Aldrich |
Cat# D2650 |
DMEM with 4.5 g/L glucose, L-glutamine & sodium pyruvate |
Cellgro |
Cat# 10–013-CV |
DNA Polymerase I, Large (Klenow) Fragment |
New England Biolabs |
Cat# M0210L |
EZview™ Red Anti-HA Affinity Gel |
Sigma-Aldrich |
Cat# E6779-1ML |
Geneticin Selective Antibiotic (G-418 Sulfate) |
Thermo Fisher Scientific |
Cat# 10131035 |
Geneticin Selective Antibiotic (G418 Sulfate) |
Thermo Fisher Scientific |
Cat# 10131035 |
HEPES |
Life Technologies |
Cat# 15630-130 |
ImmPACT DAB peroxidase (HRP) substrate |
Vector |
Cat# SK-4105 |
Isopropanol |
Sigma-Aldrich |
Cat# 190674 |
Klenow Fragment (3′-5′ exo-) |
New England Biolabs |
Cat# M0212L |
Opti-MEM® |
Thermo Fisher Scientific |
Cat# 31985062 |
Penicillin/Streptomycin |
Thermo Fisher Scientific |
Cat# 15140122 |
Phusion Flash High-Fidelity PCR Master Mix |
Thermo Fisher Scientific |
Cat# F548S |
Phusion® Hot Start Flex DNA Polymerase |
New England Biolabs |
Cat# M0535 |
Pierce™ Anti-HA Magnetic Beads |
Thermo Fisher Scientific |
Cat# 88837 |
Polybrene |
EMD Millipore |
Cat# TR-1003-G |
Polyethylenimine, Linear, MW 25,000 (PEI 25000) |
Polysciences |
Cat# 23966-1 |
Puromycin dihydrochloride |
Sigma-Aldrich |
Cat# P8833 |
RPMI 1640 w/ L-glutamine |
Fisher Scientific |
Cat# MT10040CV |
T4 DNA polymerase |
New England Biolabs |
Cat# M0203L |
T4 polynucleotide kinase |
New England Biolabs |
Cat# M0201L |
TRIzol Reagent |
Thermo Fisher Scientific |
Cat# 15596018 |
Critical commercial assays |
Agilent High Sensitivity DNA Kit |
Agilent Genomics |
Cat# 5067-4626 |
FITC Annexin V Apoptosis Detection Kit |
BD Biosciences |
Cat# 556547 |
FITC BrdU Flow Kit |
BD Biosciences |
Cat# 559619 |
Gibson Assembly Master Mix |
New England Biolabs |
Cat# E2611 |
In-Fusion HD Cloning Kit |
Takara Bio |
Cat# 638909 |
MiniElute PCR Purification Kit |
QIAGEN |
Cat# 28004 |
MiSeq Reagent Kit V3 150-cycle |
Illumina |
Cat# MS-102-3001 |
Pierce™ BCA Protein Assay Kit |
Thermo Fisher Scientific |
Cat# 23225 |
Power SYBR Green Master Mix |
Thermo Fisher Scientific |
Cat# 4368577 |
QIAamp DNA Mini Kit |
QIAGEN |
Cat# 51304 |
QIAquick Gel Extraction Kit |
QIAGEN |
Cat# 28704 |
Quick Ligation Kit |
New England Biolabs |
Cat# M2200L |
TruSeq RNA Sample Prep Kit v2 |
Illumina |
Cat# RS-122-2001 |
Deposited data |
RNA-Seq |
This study |
GEO: GSE174169
|
RNA-Seq |
Tarumoto et al. (2018)
|
GEO: GSE109491
|
RNA-Seq |
Xu etal. (2018)
|
GEO: GSE104308
|
Human reference genome GRCh37/hg19 |
Genome Reference Consortium |
https://www.ncbi.nlm.nih.gov/grc/human
|
Human reference genome GRCh37/hg38 |
Genome Reference Consortium |
https://www.ncbi.nlm.nih.gov/grc/human
|
Raw Mass Spectrometry Data Files |
This study |
ProteomeXchange Consortium with the dataset identifier PXD029769. Additional identifiers https://doi.org/10.25345/C5786Q, MassIVE (MSV000088383) |
Raw Metabolomics Data Files |
This study |
MetaboLights database with identifier MTBLS3707 |
Experimental models: Cell lines |
Human: A549 |
ATCC |
CCL-185 |
Human: AsPC-1 |
ATCC |
CRL-1682 |
Human: HEK293T |
ATCC |
CRL-3216 |
Human: HEL |
ATCC |
TIB-180 |
Human: K562 |
ATCC |
CCL-243 |
Human: Kasumi-1/Cas9 |
Robert Roeder |
N/A |
Human: MA9-ITD |
Wunderlich et al. (2013)
|
N/A |
Human: MA9-RAS |
Wunderlich et al. (2013)
|
N/A |
Human: MIAPaCa-2 |
ATCC |
CRL-142 |
Human: ML-2 |
DSMZ |
ACC 15 |
Human: MOLM-13 |
DSMZ |
ACC 554 |
Human: MV4-11 |
ATCC |
CRL-9591 |
Human: NCI-H1048 [H1048] |
ATCC |
CRL-5853 |
Human: NOMO-1 |
DSMZ |
ACC 542 |
Human: OCI-AML3 |
DSMZ |
ACC 582 |
Human: PANC-1 |
ATCC |
CRL-1469 |
Human: RD |
ATCC |
CCL-136 |
Human: RH30 |
DSMZ |
ACC 489 |
Human: RH4 |
NA |
RRID: CVCL_5916 |
Human: SEM |
DSMZ |
ACC 546. |
Human: SET-2 |
DSMZ |
ACC 608 |
Human: SUIT-2 |
JCRB |
JCRB1094 |
Human: THP-1 |
ATCC |
TIB-202 |
Human: U937 |
ATCC |
CRL-1593.2 |
Mouse: NIH3T3 |
ATCC |
CRL-1658 |
Mouse: RN2 |
Zuber etal. (2011)
|
N/A |
Experimental models: Organisms/strains |
Mouse: NSG |
The Jackson Laboratory |
Stock No: 005557 |
Oligonucleotides |
sgRNA sequences, see Tables S3 and S4
|
This study |
N/A |
Primers, see Tables S3 and S4
|
This study |
N/A |
Recombinant DNA |
LRG (Lenti_sgRNA_EFS_GFP) |
Shi etal. (2015)
|
Addgene plasmid # 65656 |
LRG2.1 |
Tarumoto et al. (2018)
|
Addgene plasmid # 108098 |
LRCherry2.1 |
Tarumoto et al. (2018)
|
N/A |
LRG2.1-Blast |
Klingbeil, unpublished |
N/A |
LentiV_Cas9_puro |
Tarumoto et al. (2018)
|
Addgene plasmid # 108100 |
LentiV_Neo |
Tarumoto et al. (2018)
|
N/A |
Lenti-luciferase-P2A-Neo |
Wei etal. (2021)
|
Addgene plasmid # 105621 |
psPAX2 |
N/A |
Addgene plasmid # 12260 |
CTDSPL2 cDNA |
Horizon Discovery |
Clone ID: 5744745 |
partial STK35 cDNA |
Horizon Discovery |
Clone ID: 9021751 |
PDIK1L cDNA |
Horizon Discovery |
Clone ID: 4828997 |
Software and algorithms |
Aminode |
Chang et al. (2018)
|
http://www.aminode.org/
|
CSHL sgRNA Design Tool |
El Demerdash, unpublished |
https://mojica.cshl.edu
|
DESeq2 |
Love et al. (2014)
|
https://bioconductor.org/packages/release/bioc/html/DESeq2.html
|
FlowJo |
TreeStar Inc. |
https://www.flowjo.com
|
GraphPad Prism v.5 and v.9 |
GraphPad Software |
https://www.graphpad.com/scientific-software/prism/
|
HTSeq-Count v.0.6.1p1 |
Anders et al. (2015)
|
https://htseq.readthedocs.io/en/master/
|
HUGO Gene Nomenclature Committee (HGNC) |
N/A |
https://www.genenames.org
|
IUPred2A |
Meszaros et al. (2018)
|
https://iupred2a.elte.hu
|
Kallisto |
Bray et al. (2016)
|
https://pachterlab.github.io/kallisto/
|
Metascape |
Tripathi et al. (2015)
|
http://metascape.org
|
Salmon v.1.0 |
Patro et al. (2017)
|
https://salmon-tddft.jp/wiki/Download-v.1.0.0
|
STAR v.2.5.2b-1 |
Dobin et al. (2013)
|
https://github.com/alexdobin/STAR
|
UCSC Genome Browser |
UCSC |
http://genome.ucsc.edu
|