HEp-2 (ATCC CCL-23) and A549 cells (ATCC CCL-185) were purchased from the American Type Culture Collection. A549-Dual, A549-Dual KO-RIG-I, A549-Dual KO-MDA5, and A549-Dual KO-MAVS-knockout cells were purchased from InvivoGen (San Diego, CA), and were supplemented with Normocin (100 μg ml−1), blasticidin (10 μg ml−1) and zeocin (100 μg ml−1). The wild type U2OS cells and METTL3-knockdown U2OS cell lines were generous gifts from Dr. Yang Shi (Harvard Medical School, Boston, MA). All cell lines were grown in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies, Carlsbad, CA) supplemented with 10% FBS and were confirmed free from mycoplasma by the LookOut Mycoplasma PCR Detection Kit (Sigma, St. Louis, MO). Recombinant RSV (rgRSV) strain A2 expressed GFP as the first gene, and rgRSV-ΔG was derived from rgRSV by deletion of the G gene [76 (link),77 (link)].
A549 dual ko rig 1
Manufactured by InvivoGen
The A549-Dual™ KO-RIG-I is a human alveolar basal epithelial cell line. It is genetically engineered to lack the RIG-I (retinoic acid-inducible gene I) receptor, which is a key sensor of viral RNA. This cell line can be used for the study of the innate immune response to viral infections.
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Lab products found in correlation
A549 dual, by InvivoGen (5 mentions)
A549 dual ko mda5, by InvivoGen (4 mentions)
Lookout mycoplasma pcr detection kit, by Merck Group (3 mentions)
A549 dual ko mavs knockout cells, by InvivoGen (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Puromycin, by Thermo Fisher Scientific (2 mentions)
Thp 1 atcc tib 202, by American Type Culture Collection (2 mentions)
A549 cells atcc ccl 185, by American Type Culture Collection (1 mentions)
Hep 2 atcc ccl 23, by American Type Culture Collection (1 mentions)
Vero e6 cell, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Trypan blue, by Thermo Fisher Scientific (1 mentions)
Countess 2, by Thermo Fisher Scientific (1 mentions)
Accutase solution, by Thermo Fisher Scientific (1 mentions)
Zeocin, by InvivoGen (1 mentions)
Blasticidin, by InvivoGen (1 mentions)
Celltiter glo luminescent cell viability assay, by Promega (1 mentions)
Thp1 dual, by InvivoGen (1 mentions)
Hek lucia rig 1, by InvivoGen (1 mentions)
6 protocols using a549 dual ko rig 1
1
Respiratory Syncytial Virus Cell Lines
2
Establishing Cell Lines for Virus Research
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Vero E6 cells (ATCC CRL-1586), A549 cells (ATCC CCL-185), and THP-1 (ATCC TIB-202) were purchased from the American Type Culture Collection (Manassas, VA). A549-Dual™, A549-Dual™ KO-RIG-I, A549-Dual™ KO-MDA5, and A549-Dual™ KO-MAVS knockout cells were purchased from InvivoGen (San Diego, CA). BHK-SR19-T7 cells were kindly provided by Apath, LLC, Brooklyn, NY. All cell lines were grown in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies) supplemented with 10% FBS. The medium for the BHK-SR19-T7 cells was supplemented with 10 μg/ml puromycin (Life Technologies) during every other passage to select for T7 polymerase-expressing cells. A549-Dual™ and knockout cell lines were supplemented with Normocin™ (100 μg/ml), blasticidin (10 μg/ml) and Zeocin™ (100 μg/ml). HeLa cells overexpressing the empty vector (pPB-CAG), YTHDF1, YTHDF2, or YTHDF3 were maintained in DMEM supplemented with 10% FBS and 1 µg/ml of puromycin every passage to select for YTHDF1-3 overexpressing cells. All cell lines used in this study were free of mycoplasma, as confirmed by the LookOut Mycoplasma PCR Detection Kit (Sigma). Cell lines were authenticated by the ATCC or InvivoGen.
3
Establishing Cell Lines for Virus Research
4
Cell Culture Protocol for COVID-19 Research
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A549 cells (ATCC; CCL-185), A549ACE2 cells (67 (link)), HEK-293T cells (ATCC; CRL-3216), HEK-293T-NoDice cells (68 (link)), HeLa cells (ATCC; CCL-2), Vero E6 cells (ATCC; CRL-1586), BHK-21 cells (ATCC; CCL-10), and MDCK cells (ATCC; CCL-34) were all maintained in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS). A549-Dual, A549-Dual-KO-MAVS, A549-Dual-KO-MDA5, and A549-Dual-KO-RIG-I cells were acquired from InvivoGen and maintained in DMEM supplemented with 10% FBS, 100 μg/ml Normocin, 10 μg/ml blasticidin, and 100 μg/ml zeocin. All cells were cultured at 37°C and 5% CO2.
5
Culturing A549 Dual-reporter Cell Lines
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A549-Dual and A549-Dual RIG-I KO (InvivoGen, Ref. a549d-nfis or a549d-korigi, San Diego, California, U.S.A.) were cultured with high glucose Dulbecco’s Modified Eagles Medium containing 1% GlutaMAX™ (DMEM, Gibco™, ThermoFisher, Ref. 10566016) and supplemented with 10 μg/mL Blasticidin (InvivoGen, Ref. ant-bl-05), 100 μg/mL Zeocin (InvivoGen, Ref. ant-zn-05), 10% v/v heat inactivated Fetal Bovine Serum (FBS, Gibco™, ThermoFisher, Ref. 10099141). Proliferative cultures were incubated at 37°C in a humidified 5% CO2 incubator and subculture carried out twice a week by washing the cell monolayers with DPBS followed by addition of accutase solution (Gibco™, ThermoFisher, Ref. A1110501) and incubation at 37°C until the cells detached. Accutase was inactivated by the addition of growth medium before seeding 1.0.106 cells into fresh T175 or T75 flasks. Cell numbers and viability were assessed by Trypan Blue staining (Thermo Fisher Scientific, Ref. T10282) using Countess 2 (Invitrogen, Ref. AMQAX1000).
6
Murine Cell Lines and Reporter Assays
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B16F10, CT26, 4T1, and MC38 murine lines were purchased from American Type Culture Collection, expanded, confirmed to be mycoplasma free, and then frozen as aliquots. HEK reporter lines, A549-Dual, and the derived genetic KO reporter lines were purchased from Invivogen: HEK-Lucia Null (hkl-null), HEK-Lucia RIG-I (hkl-hrigi), A549-Dual (a549d-nfis), A549-Dual-RIG-I-KO (a549d-korigi), A549-Dual-MDA5-KO (a549d-komda5), A549-Dual-MAVS-KO (a549d-komavs), THP1-Dual (thpd-nfis), THP1-Dual-IRF3-KO (thpd-koirf3), and THP1-Dual-MAVS-KO (thpd-komavs). All reporter cell lines from Invivogen were maintained as recommended by the manufacturer. Murine cell lines were maintained in American Type Culture Collection recommended media. Viability assays were performed using the CellTiter-Glo Luminescent Cell Viability Assay (Promega; G7573) following the manufacturer's instructions.
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