Gene pairs associated with a synthetic lethal phenotype in the library screen were validated using combinatorial viability screening of sgRNA perturbations with 5 small molecule inhibitors: navitoclax (ABT-263, Active Biochem A-1001), A1331852 (Active Biochem A-6048); venetoclax (ABT-199, Active Biochem A-1231); WEHI539 (MedChem Express, HY-15607A); and the MCL1 inhibitor S63845 (a gift from Guo Wei, Golub lab). Meljuso cells were transduced in 12-well plates, as described above, with lentivirus containing a single sgRNA targeting one of the anti-apoptotic genes (BCL2L1, BCL2L2, MCL1) or a control sgRNA either targeting CD81 or containing a run of 6 thymidines. Two days after transduction, cells were selected using puromycin at 1 μg/mL for five days. After puromycin selection, 3,000 cells were seeded into 96-well plates. Across each row of the 96-well plate, a different small molecule was added at 11 log2-dilutions ranging from 1 μM to approximately 1 nM in duplicate for each of the cell lines. The last well in the row did not receive small molecule. After 3 days in the presence of the small molecule, viability of the cell population was assayed by CellTiterGlo (Promega) according to the manufacturer’s instructions.
A1331852
Manufactured by Active Motif
The A1331852 is a laboratory equipment product. It functions as a core component in various experimental setups. The product specifications and technical details are not readily available without access to the full product information.
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Lab products found in correlation
2 protocols using a1331852
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Synthetic Lethal Validation via Combinatorial Screening
2
Synthetic Lethal Validation via Combinatorial Screening
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Gene pairs associated with a synthetic lethal phenotype in the library screen were validated using combinatorial viability screening of sgRNA perturbations with 5 small molecule inhibitors: navitoclax (ABT-263, Active Biochem A-1001), A1331852 (Active Biochem A-6048); venetoclax (ABT-199, Active Biochem A-1231); WEHI539 (MedChem Express, HY-15607A); and the MCL1 inhibitor S63845 (a gift from Guo Wei, Golub lab). Meljuso cells were transduced in 12-well plates, as described above, with lentivirus containing a single sgRNA targeting one of the anti-apoptotic genes (BCL2L1, BCL2L2, MCL1) or a control sgRNA either targeting CD81 or containing a run of 6 thymidines. Two days after transduction, cells were selected using puromycin at 1 μg/mL for five days. After puromycin selection, 3,000 cells were seeded into 96-well plates. Across each row of the 96-well plate, a different small molecule was added at 11 log2-dilutions ranging from 1 μM to approximately 1 nM in duplicate for each of the cell lines. The last well in the row did not receive small molecule. After 3 days in the presence of the small molecule, viability of the cell population was assayed by CellTiterGlo (Promega) according to the manufacturer’s instructions.
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