Key resources table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit anti-gamma H2A.X (phospho S139) | Abcam | Cat#ab81299 |
| Anti-alpha-tubulin-FITC antibody | Sigma | Cat#F2168 |
| Goat anti-rabbit antibody disulfide-linked
to Alexa Fluor 594 (custom conjugation) |
Invitrogen; Thermo Fisher Scientific |
Cat#21212; Cat#A10270 |
| Rabbit anti-GFP | Cheeseman Lab | N/A |
| Mouse anti-gamma H2A.X (phospho S139) | Millipore | Cat#05636 |
| Rabbit anti c-Myc | Abcam | Cat#ab32072 |
| Rabbit anti-“Bonsai”/NDC80 | Schmidt et al., 201293 | N/A |
| HRP Anti-GAPDH antibody | Abcam | Cat#Ab185059 |
| Mouse Anti-alpha-tubulin | Sigma | Cat#T6199 |
| Rabbit anti-centrin | Backer et al., 201294 | N/A |
| Mouse anti-CENPA | Invitrogen | Cat#MA1–20832 |
| Chemicals, peptides, and recombinant proteins | ||
| Alexa Fluor Plus 750 Phalloidin | Thermo Fisher Scientific | Cat#A30105 |
| Critical commercial assays | ||
| RevertAid H minus Reverse Transcriptase | Thermo Fisher Scientific | Cat#EP0452 |
| Ribolock RNase inhibitor | Thermo Fisher Scientific | Cat#EO0384 |
| RNase H | Enzymatics | Cat#Y9220L |
| TaqIT DNA polymerase | Enzymatics | Cat#P7620L |
| Ampligase | Lucigen | Cat#A3210K |
| Phi29 DNA polymerase | Thermo Fisher Scientific | Cat#EP0091 |
| MiSeq Reagent Nano Kit v2 with PR2 buffer | Illumina | Cat#MS-103–1003 |
| MitoTracker Orange CMTMRos | Invitrogen | Cat#M7510 |
| Deposited data | ||
| Processed images and data | This paper | https://vesuvius.wi.mit.edu |
| Processed single-cell phenotype measurements | This paper | Harvard Dataverse https://doi.org/10.7910/DVN/VYKTI5 |
| Raw image data | This paper | BioImage Archive S-BIAD394 |
| RNA-sequencing data | This paper | Gene Expression Omnibus: GSE199901 |
| Experimental Models: Cell lines | ||
| All cell lines used in this study, see Table S5 | This study | N/A |
| Oligonucleotides | ||
| All oligonucleotides used in this study, see Table S1 | This study | N/A |
| Recombinant DNA | ||
| CROPseq-puro-v2 | Feldman et al., 20198 | #127458 (Addgene) |
| sgOPTI | Fulco et al., 201688 | #85681(Addgene) |
| pBABE H2B-mCherry (pKC96) | This paper | Undergoing submission (Addgene) |
| pBABE GFP-SPC24 (pKG422) | Gascoigne et al., 201187 | Undergoing submission (Addgene) |
| pBABE EGFP-LIN52 (pKC518) | This paper | Undergoing submission (Addgene) |
| pBABE EGFP-ZNF335 (pKC530) | This paper | Undergoing submission (Addgene) |
| Software and algorithms | ||
| General repository used for processing optical pooled screening data | Feldman et al., 202215 |
https://github.com/feldman4/OpticalPooledScreens (https://doi.org/10.5281/zenodo.5002684) |
| Application-specific tools for the screens presented in this paper | This paper |
https://github.com/lukebfunk/OpticalPooledScreens (https://doi.org/10.5281/zenodo.7155628) |