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. 2020 Nov 21;11(11):998. doi: 10.1038/s41419-020-03196-0

Table 2.

Listing/summary of model generation strategies using overexpression of IDHMUT on wild-type background.

Type of cells Method Vector Protein or metabolite validation Results of IDH1 mutation References
Overexpression: I. Transduction
IMA Lentivirus pLenti6.3/TO/V5-EST MS Suppress anchorage-independent growth 55
U87MG and U251 Lentivirus pLenti6.3-MCS-IRES2-EGFP WB Reduces proliferation, cell survival, and invasion of human glioma by downregulating Wnt/-catenin signaling 51
U87MG and U373MG Retrovirus pLPCX vector MS Sensitivity to radiation therapy 11
hESC-NSC Lentivirus pEF1α-IDH1/pEF1α-R132H MS IDH1 combined with P53, and ATRX mutation block differentiation 31
hiPSC-NPC (ebiNSc) Lentivirus pLEX_307 plasmid gateway tec WB, ICC Sensitivity to radiation 61
IMA cell (IDH1 mutated astrocytoma) Lentivirus pLenti6.2/V5 MS IDH1 mutation produce D2HG 18
GP2-293 cells Retrovirus pCMV-Sport6 MS Blocking differentiation 96
Immortalized human astrocytes Lentivirus pLNCX2 MS Hypermethylation 62
NHA-hTERT. HeLa, HT1080 and 293E Retrovirus pLPCX MS mTOR activation 66
U87, NHA Lentivirus – WB Increases RAD51-mediated HR and TMZ resistance 97
U87, HCT116, BT-142, T98G, LN229 Retrovirus pLPCX WB Lower level of MCI-1 protein 98
U87 and U251 Lentivirus PGC-FU-GFP – Decreasing GSH and increasing ROS 99
Mesenchymal stem cell Lentivirus pDONR221 MS Dysregulate differentiation 100
SF10602, BT-142, GM847, MGG119 Lentivirus GFP-tagged RAP1 or RFP-tagged XRCC1 WB Telomeric dysfunction and alters DNA repair 101
U87 Lentivirus U87, NHA WB Inhibition PIK3mTOR and reduction D2HG 102
II. Transfection
U87 Transfection p-EGFP-C1 WB Enhancement of chemotherapy Sensitivity 103
U87 Lipofectamin 3000 pCMV-GV23 WB Increased levels of repressive H3K9me3, enhancement of chemotherapy Sensitivity 73
SVGp12 and U251 Lipofectamin pCMV-Sport6 WB Induced proliferation of glioma cells via NFkB in a HIF1-α manner 104
293T, U87MG, LN-18 Lipofectamine 2000 pCMV-6 MS Mutated IDH1 produce D2HG 20
U87, U138 Geneticin p-EGFP-C2-bio MS Decreasing proliferation of IDH1-R132H 33
U87MG Lipofectamine 2000 pCMV-Tag2B WB Upregulating AKT-mTOR 36
LN229 Fugene 6 – Enzymatic assay Neo-enzymatic activity for D2HG production 23
Murine GL261, SB28 Lipofectamine 2000 – MS Suppress STAT1 and CD8+ T cell accumulation 46
HT1080 Lipofectamine 2000 pCMV-Sport6 MS Oncometabolite production 105
U251, Hela Calcium phosphate pcDNA6 WB Acetate accumulation 106