Simettl3 1

Manufactured by GenePharma

Sourced in China

SiMETTL3#1 is a laboratory equipment designed for gene expression analysis. It utilizes the CRISPR-based gene editing technology to modify target genes. The core function of SiMETTL3#1 is to enable precise and efficient genetic manipulations in research settings.

Automatically generated - may contain errors

Lab products found in correlation

Simettl3 2, by GenePharma (4 mentions) Si ythdf1, by GenePharma (2 mentions) Pex 3 mettl3 expression plasmid, by GenePharma (2 mentions) Pcdna3.1 slc7a11 expression plasmid, by Genomed (2 mentions) Sh mettl3, by GenePharma (2 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Si socs2, by Santa Cruz Biotechnology (1 mentions) Oil red o, by Merck Group (1 mentions) Si nc, by GenePharma (1 mentions) Ox ldl, by Biomedical Technologies (1 mentions)

4 protocols using simettl3 1

Targeted Knockdown and Overexpression of METTL3

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The siRNAs for METTL3 and YTHDF1, and lentivirus for METTL3 knockdown were synthesized by GenePharma (Shanghai, China). The sequences were as follows: siMETTL3#1 (sense: 5′-GCUACCUGGACGUCAGUAUTT-3′, antisense: 5′-AUACUGACGUCCAGGUAGCTT-3′); siMETTL3#2 (sense: 5′-GGUUGGUGUCAAAGGAAAUTT-3′, antisense: 5′-AUUUCCUUUGACACCAACCTT-3′); siYTHDF1 (sense: 5′-GGAGAAUAACGACAACAAATT-3′, antisense: 5′-UUUGUUGUCGUUAUUCUCCTT-3′); and shMETTL3 (5′-GCAAGAATTCTGTGACTATGG-3′).
The pEX-3-METTL3 expression plasmid was synthesized by GenePharma (Shanghai, China). The pcDNA3.1-SLC7A11 expression plasmid was synthesized by Genomeditech (Shanghai, China).

Xu Y., Lv D., Yan C., Su H., Zhang X., Shi Y, & Ying K. (2022). METTL3 promotes lung adenocarcinoma tumor growth and inhibits ferroptosis by stabilizing SLC7A11 m6A modification. Cancer Cell International, 22, 11.

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Transient METTL3 Knockdown in Cells

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To transiently knock down METTL3 expression, siRNA transfection was used. Two siRNAs targeting METTL3 (si-METTL3#1 and si-METTL3#2) were synthesized by GenePharma Corporation. The siRNA sequences were as follows: si-METTL3#1, 5′-GGUGACUGCUCUUUCCUUATT-3′ and 3′-UAAGGAAAGAGCAGUCACCTT-5′; si-METTL3#2, 5′-GCUACCUGGACGUCAGUAUTT-3′ and 5′-AUACUGACGUCCAGGUAGCTT-3′. The negative control siRNA sequences were as follows: si-Ctrl, 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′. The siRNA targeting SOCS2 (si-SOCS2) was obtained from Santa Cruz Biotechnology, Inc. The pCMV6 plasmid containing the full-length SOCS2 sequence (FULL-SOCS2) was purchased from Origene (RC203163).
Cells were cultured in 6-well plates (1×10⁵ cells per well) overnight prior to the experiment. The following day, siRNA (80 µM) was transfected into cells with Lipofectamine RNAiMAX (5 µl per well) (Thermo Fisher Scientific, Inc.). For plasmid transfection, cells were seeded at a density of 1×10⁵ cells per well in 6-well plates. Twenty-four hours later, Lipofectamine 2000 (5 µl per well; Thermo Fisher Scientific, Inc.) was used to transfect cells with the plasmid (2.5 µg per well) following the manufacturer's instructions. Cells were harvested 48 h after transfection for RNA and protein detection.

Xu J., Chen Q., Tian K., Liang R., Chen T., Gong A., Mathy N.W., Yu T, & Chen X. (2020). m6A methyltransferase METTL3 maintains colon cancer tumorigenicity by suppressing SOCS2 to promote cell proliferation. Oncology Reports, 44(3), 973-986.

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Regulation of Endothelial Cell Function by IGF2BP1 and METTL3

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Ox-LDL was obtained from Biomedical Technologies, Inc. (Stoughton, MA, United States). Oil red O was obtained from Sigma-Aldrich, Inc. (St. Louis, MO, United States). Small interference RNAs (siRNA) targeting IGF2BP1 (si-IGF2BP1#1, si-IGF2BP1#2, and si-IGF2BP1#3), METTL3 (si-METTL3#1, si-METTL3#2, and si-METTL3#3), and a scrambled siRNA (si-NC) were obtained from GenePharma Co., Ltd. (Shanghai, China). The siRNA sequences were shown in Table 1. The recombinant plasmid expressing JAK2 (pcDNA-JAK2) or IGF2BP1 (pcDNA-IGF2BP1) was synthesized by GeneCreate Biological Engineering Co., Ltd. Plasmids or siRNAs were transfected into HUVECs using jetPRIME transfection reagent (Poluplus-transfection Inc., New York, NY, United States) according to the instructions of the manufacturer. For double transfection, DNA (2 μg) and siRNA (final concentration: 50 nM) were mixed and co-incubated with 5 μl jetPRIME reagent for 15 min at room temperature in 6-well plates. Next, the transfection mix was added to cells in serum-containing medium dropwise. At 24 h after transfection, the transfection medium was replaced with the refresh cell growth medium.

Dong G., Yu J., Shan G., Su L., Yu N, & Yang S. (2021). N6-Methyladenosine Methyltransferase METTL3 Promotes Angiogenesis and Atherosclerosis by Upregulating the JAK2/STAT3 Pathway via m6A Reader IGF2BP1. Frontiers in Cell and Developmental Biology, 9, 731810.

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METTL3 and YTHDF1 RNA Silencing Constructs

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The siRNAs for METTL3 and YTHDF1, and lentivirus for METTL3 knockdown were synthesized by GenePharma (Shanghai, China). The sequences were as follows:
siMETTL3#1 (sense: 5′-GCUACCUGGACGUCAGUAUTT-3′, antisense: 5′-AUACUGACGUCCAGGUAGCTT-3′); siMETTL3#2 (sense: 5′-GGUUGGUGUCAAAGGAAAUTT-3′, antisense: 5′-AUUUCCUUUGACACCAACCTT-3′); siYTHDF1 (sense: 5′-GGAGAAUAACGACAACAAATT-3′, antisense: 5′-UUUGUUGUCGUUAUUCUCCTT-3′); and shMETTL3 (5'-GCAAGAATTCTGTGACTATGG-3').
The pEX-3-METTL3 expression plasmid was synthesized by GenePharma (Shanghai, China). The pcDNA3.1-SLC7A11 expression plasmid was synthesized by Genomeditech (Shanghai, China).

Xu Y., Lv D., Yan C., Su H., Zhang X., Shi Y., & Ying K. (2021). METTL3 Promotes Lung Adenocarcinoma Tumorigenesis and Inhibits Ferroptosis by Stabilizing SLC7A11 m6A Modification.

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