REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
α-INTS10 | Abcam | Cat# ab180934s |
α-INTS13 | Bethyl | Cat# A303-575A; RRID: AB_11125549 |
α-INTS14 | Prestige | Cat# HPA040651 |
α-C7orf26 | Prestige | Cat# HPA052175 |
α-His | CST | Cat# 2366 |
α-Mouse | Licor | Cat# 926-32210 |
α-Rabbit | Licor | Cat# 926-32213 |
α-CD11b AF647 | Biolegend | Cat# 101220; RRID: AB_493546 |
Bacterial and virus strains | ||
MegaX Competent Cells | ThermoFisher | Cat# C640003 |
Stellar Competent Cells | Takara | Cat# 636766 |
Chemicals, peptides, and recombinant proteins | ||
TransIT-LT1 Transfection Reagent | Mirus Bio | Cat# MIR2300 |
Critical commercial assays | ||
Chromium Single-Cell 3ʹ v3 with Feature Barcoding | 10x Genomics | PN-1000075, PN-1000153, PN-1000079 |
Seahorse XF Cell Mito Stress Test | Agilent | Cat# 103015-100 |
Bioanalyzer RNA nano | Agilent | Cat# 5067-1511 |
Deposited data | ||
Raw sequencing data from Perturb-seq screens | This paper | SRA BioProject PRJNA831566 |
Processed data from Perturb-seq screens | This paper | http://gwps.wi.mit.edu |
Experimental models: Cell lines | ||
K562 dCas9-BFP-KRAB | Gilbert et al., 2014 | N/A |
RPE1 Zim3-dCas9 | This study | N/A |
Oligonucleotides | ||
Dual sgRNA gDNA primer: AATGATACGGCGACCACCGAGATCTACACCGCGGTCTGTATCCCTTGGAGAACCACCT | Nunéz et al., 2021 | oJR324 |
Dual sgRNA gDNA index primer: CAAGCAGAAGACGGCATACGAGATnnnnnGCGGCCGGCTGTTTCCAGCTTAGCTCTTAAA | Nunéz et al., 2021 | oJR325 |
Custom R1 sequencing primer: CGCGGTCTGTATCCCTTGGAGAACCACCTTGTTGG | Nunéz et al., 2021 | oJR326 |
Custom R2 sequencing primer: GCGGCCGGCTGTTTCCAGCTTAGCTCTTAAAC | Nunéz et al., 2021 | oJR328 |
Custom Index Read 1 sequencing primer: GTTTAAGAGCTAAGCTGGAAACAGCCGGCCGC | Nunéz et al., 2021 | oJR327 |
Recombinant DNA | ||
See Table S9 for plasmids used in this study | This paper | N/A |
Software and Algorithms | ||
CellRanger 4.0.0 | 10X Genomics, Inc. | http://software.10xgenomics.com |
sgRNA assignment scripts | Replogle et al., 2020 | https://github.com/josephreplogle/guide_calling |
Perturb-seq analysis codebase | Norman et al., 2019 | https://github.com/thomasmaxwellnorman/Perturbseq_GI |