Table 2.
KRAS synthetic lethal CRISPR-based functional screens.
| Reference | CRISPR Library | Type of Study | Cell Lines | Combined Screen | Synthetic Lethal Hits or Pathways |
|---|---|---|---|---|---|
| Wang et al. Cell. 2017 [115] | Genome-wide (GW) human CRISPR libraries | Pooled CRISPR-based screen (proliferation assay) | 6 human KRAS/NRAS-mutant and 6 KRAS-WT leukemia cell lines | Parallel GW mouse CRISPR screen using isogenic Ba/F3 cells (NRAS) | RCE1, ICMT, RAF1, SHOC2, PREX1 |
| Martin et al. Cell Rep. 2017 [117] | Genome-wide CRISPR library targeting 18,148 genes | Pooled CRISPR-based screen (proliferation assay) | Isogenic pairs of HCT116/DLD1 cells (KRASG13D); LS513 cells (KRASG12D). Colorectal cancer cells | shRNA library targeting 1100 essential genes | Mitochondrial protein translation, transcription and oxidative phosphorylation pathways; Mrpl52 and Ndufb10 |
| Anderson et al. Cell Rep. 2017 [86] | Custom CRISPR/Cas9 library (~2000 sgRNAs) targeting 378 genes | Pooled CRISPR-based screen (drug sensitizer screening) | Pan-cancer panel of KRAS-mutant cell lines | Drug inhibition of KRAS pathways: MEK, ERK, PI3K. | MAPK14, MDM4, SRC |
| Yau et al. Cancer Res. 2017 [118] | Human GeGKO v2 library pooled plasmid (lentiCRISPRv2) | Pooled CRISPR-based in vivo screen (xenograft model) | Isogenic pairs of HCT116 cells (KRASG13D). Colorectal cancer cells | Secondary smaller focused CRISPR screen targeting ~320 KRAS-related genes | NADK, KHK, SUCLA2, INO80C. Nucleotide synthesis, redox balance and mitochondrial processes |
| Šuštić et al. Genome Med. 2018 [119] | Human GeGKO v2 library pooled plasmid (lentiCRISPRv2) | Pooled CRISPR/Cas9 MEK inhibitor resistance screen | ERN1KO LoVo cells (KRASG13D). Colorectal cancer cells | MEK inhibitors: Selumetinib and Trametinib | DUSP4, STK40, RUNX2, CBFB, DET1, COP1. Negative regulation of the JUN signaling |
| Szlachta et al. Nat Commun. 2018 [120] | CRISPR library from Dr. Sabatini (~4000 human genes) | Pooled CRISPR knockout sensitizer screen | PDX366 model (KRAS, P53 and SMAD4 mutant). Pancreatic PDX-cells | MEKi: Trametinib | CENPE, RRM1 |
| Dompe et al. PLoS ONE. 2018 [121] | Custom druggable genome CRISPR library (2194 genes) | Pooled CRISPR knockout sensitizer screen | MOR lung cancer cell line (KRAS-mutant) | MEKi: Cobimetinib and ERKi: GDG-0994. +Validation focused screens (4 KRAS-mutant lung cancer cells) |
MAPK7 |
| Sulahian et al. Cell Rep. 2019 [122] | Genome scale Avana-4 barcoded CRISPR library (74,687 sgRNAs) | Pooled CRISPR-Cas9 screens MEK sensitizer screen | KRAS-mutant cancer cell lines (pancreas and lung) | MEKi: Trametinib. | SHOC2, BCL2L1, MCL1, EXT1, EXT2, EXTL3, SLC35B2. |
| Han et al. Nature. 2020 [123] | Genome-Wide custom CRISPR library | 2D vs. 3D Pooled CRISPR-Cas9 screen (proliferation assays) | H23 KRAS-mutant cells (KRASG12C). Lung adenocarcinoma (LUAD) | KRASi: ARS-853 | CPD, IGF1R |
| Michels et al. Cell Stem Cell. 2020 [124] | Custom CRISPR library (85 tumor suppressor genes) | In vivo screen (tumor growth study) | Colon organoids (APC−/−/KRASG12D) vs. cancer cell lines | CRISPR-UMI validation screen (281 sgRNAs) | TGFBR2 |
| Kelly et al. Cancer Discov. 2020 [125] | Custom CRISPR Double Knockout (CDKO) library (119 genes, 7021 pairs) | CDKO screen for genetic interactions (proliferation assays) | 2 KRAS-mutant cell lines (A549/H23). LUAD | Focused CDKO screen in 9 LUAD cell lines | RHOA-RAP1GDS1 combination |
| Li et al. Cancer Discov. 2020 [126,127] | Custom CRISPR library (524 epigenetic regulators) | Epigenetic-focused CRISPR KO in vivo screen | KP mutant lung cancer mouse model | Drugs: anti-PD1 or isotype control | Asf1a, Npm1 |
| Takahashi et al. Mol cell. 2020 [128] | Custom CRISPR-Cas9 library (1500 NRF2-hyperactivated related genes) | 2D vs. 3D Pooled CRISPR-Cas9 screen (proliferation assays) | A549/H1437 LUAD 2D cell lines and 3D spheroids | N/A | TSC1, GPX4 |
| Wei et al. Proc Natl Acad Sci USA. 2020 [129] | CRISPR library from Dr. Sabatini (619 human genes) | Pooled CRISPR knockout sensitizer screen | PDX366 model (KRAS, P53 and SMAD4 mutant). Pancreatic PDX-cells | Drug: Gemcitabine | PRMT5 |
| Jung et al. Oncogene. 2021 [130] | Genome-Wide CRISPR/Cas9 library | Pooled CRISPR knockout sensitizer screen | SW620 cells (KRASG12V) and HCT116 cells (KRASG13D). Colorectal cancer | Drug: ABT-263 | WNT signaling pathway; BCL-2 family genes |
| Biancur et al. Cell Metab. 2021 [131] | Custom CRISPR/Cas9 KO library (3000 mouse metabolic genes) | Pooled CRISPR KO screen: in vitro and in vivo (proliferation/viability assays) | C57BL/6 mouse PDA cell line (KRASG12D) | Additional CRISPR screen in a 3D culture model | Fdft1; cholesterol synthesis |
| Yu et al. Oncogene. 2022 [132] | Genome-Wide CRISPR/Cas9library (human GeCKO) | Pooled CRISPR knockout sensitizer screen | HCT116 cells (KRASG13D). Colorectal cancer | MEKi: AZD6244 | GRB7; RTK pathway |