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Plenti6 krasg12v

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The PLenti6-KRASG12V is a lentiviral vector that can be used to express the mutant KRAS G12V gene in target cells. The vector contains the KRAS G12V coding sequence under the control of a constitutive promoter, as well as elements necessary for lentiviral packaging and transduction.

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Lab products found in correlation

Polybrene, by Merck Group (2 mentions) Mle 12 cells, by American Type Culture Collection (2 mentions) Universal mycoplasma detection kit, by American Type Culture Collection (2 mentions) Bay11 7082, by Merck Group (1 mentions) Hbec3 kt, by American Type Culture Collection (1 mentions)

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KRASG12V (5 citations)

2 protocols using plenti6 krasg12v

1

Characterization of KRAS-driven Cell Lines

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HBEC-3KT (obtained from ATCC) and HBEC3KT-KP cells with KRASG12V expressing (/K) ± p53 shRNA downregulated (/P; a gift from Drs. John Minna and Jerry Shay; ref. 16 (link)) were cultured as described (10 (link)). HBEC3KT-KP cells were authenticated via Western immunoblotting and qPCR for p53 and KRASG12V. MLE12 cells (obtained from ATCC) were cultured as described (17 (link)). These cell lines were transduced with pLEX304-eGFP, –wild-type (WT) C9b, or -AT/GG Mut C9b lentivirus treated with 8 μg/mL polybrene (Sigma) and selected with blasticidin. MLE12/K and HBEC-3KT/K cells are generated by transducing cells with pLenti6-KRASG12V (Addgene). Plate colony formation assays (cell survival) and soft agar colony formation assays (AIG) were undertaken as described (7 (link)). All cell lines were used within 6 passages from receipt and thawing. Cell lines were tested every 2 months for Mycoplasma (Universal Mycoplasma Detection Kit, ATCC) throughout the study starting 2 months after thawing cells received from ATCC and immediately for other cell lines herein described. Parental cell lines were authenticated by short tandem repeat profiling. Cells were treated with Bay 11-7082 (Sigma, 0–10 μmol/L range) at the indicated doses.
Kim M., Vu N.T., Wang X., Bulut G.B., Wang M.H., Uram-Tuculescu C., Pillappa R., Kim S, & Chalfant C.E. (2022). Caspase 9b Drives Cellular Transformation, Lung Inflammation, and Lung Tumorigenesis. Molecular Cancer Research, 20(8), 1284-1294.
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2

KRAS and p53 Modulation in Lung Cell Lines

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HBEC-3KT (Obtained from ATCC) and HBEC3KT-KP cells with KRASG12V expressing (/K) ± p53 shRNA downregulated (/P) (a gift from Drs. John Minna and Jerry Shay (16 (link))) were cultured as described (10 (link)). HBEC3KT-KP cells were authenticated via western immunoblotting and qPCR for p53 and KRASG12V. MLE12 cells (Obtained from ATCC) were cultured as described (17 (link)). These cell lines were transduced with pLEX304-eGFP, -WT C9b, or -AT/GG Mut C9b lentivirus treated with 8 μg/mL polybrene (Sigma) and selected with blasticidin. MLE12/K and HBEC-3KT/K cells are generated by transducing cells with pLenti6-KRASG12V (addgene). Plate colony formation assays (cell survival) and soft agar colony formation assays (AIG) were undertaken as described (7 (link)). All cell lines were used within 6 passages from receipt and thawing. Cell lines were tested every two months for mycoplasma (universal mycoplasma detection kit, ATCC) throughout the study starting two months after thawing cells received from ATCC and immediately for other cell lines herein described. Parental cell lines were authenticated by STR (short tandem repeat) profiling.
Kim M., Vu N.T., Wang X., Bulut G.B., Wang M.H., Uram-Tuculescu C., Pillappa R., Kim S, & Chalfant C.E. (2022). Caspase 9b Drives Cellular Transformation, Lung Inflammation, and Lung Tumorigenesis. Molecular Cancer Research, 20(8), 1284-1294.
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