Malat1 sirna

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MALAT1 siRNA is a laboratory reagent designed to target and silence the expression of the MALAT1 gene. MALAT1 is a long non-coding RNA that has been studied for its potential role in various cellular processes. This siRNA product is intended for use in research applications to investigate the functional effects of MALAT1 knockdown.

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SiRNAs (258 citations) Malat1 (10 citations)

8 protocols using malat1 sirna

MALAT1 Knockdown and Overexpression Protocol

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The MALAT1 siRNA were designed and obtained from Invitrogen (4455877; Thermo Fisher Scientific, Inc.). The sequence of MALAT1 siRNA was 5′-GCAGAGGCAUUUCAUCCUU-3′. The sequence of the negative control siRNA was 5′-ACGUCACACGUUCGGAGAATT-3′ and it was provided by Thermo Fisher Scientific, Inc. The human NLRC5 (NCBI reference sequence, NP_115582) was obtained by nested PCR from a human cDNA library (Marathon-ready cDNA; Clontech Laboratories, Inc.) using the following primers: Forward, 5′-CGTGGGGACCCTAGAGCACCTATCA-3′ and reverse, 5′-GCATCACTTGGCTGGATTCCAAAGG-3′. The PCR products were cloned using a Takara one step PCR kit (Takara Biotechnology Co., Ltd.) and initial denaturation at 98°C for 30 sec, 30 cycles of denaturation at 98°C for 5 sec, annealing at 60°C for 10 sec, and extension at 72°C for 10 sec, finally, extension at 72°C for 2 min. For MALAT1 overexpression, the human MALAT1-expressing vector (ORF023250, NR_002819.3) was obtained from Applied Biological Materials Inc (Richmond, BC, Canada).

Liu Z., Liu J., Wei Y., Xu J., Wang Z., Wang P., Sun H., Song Z, & Liu Q. (2019). LncRNA MALAT1 prevents the protective effects of miR-125b-5p against acute myocardial infarction through positive regulation of NLRC5. Experimental and Therapeutic Medicine, 19(2), 990-998.

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MALAT1 Overexpression and Silencing in Osteosarcoma

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The procedure was carried out as previously described [63 (link)]. MNNG/HOS cells at 60~80% confluence were then selected for cell transfection. Plasmids were transfected into osteosarcoma cells by using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer's protocol. The sequence of MALAT1 siRNAs were as follows as ascribed before [21 (link), 64 (link)]: 1# MALAT1 siRNA, 5’-GGCAAUGUUUUACACUAUUTT-3’; 2# MALAT1 siRNA, 5’-CACAGGGAAAGCGAGTGGTTGGTAA-3’; 3# MALAT1 siRNA, 5’-CACAGGGAAAGCGAGUGGUUGGU-3’; non-specific siRNA 5’-UUCUCCGAACGUGUCACGUTT-3’. MALAT1-siRNA and non-specific siRNA were purchased from Invitrogen (USA). To construct MALAT1 stable over-expression cell lines for further animal study, the stable transfected pcDNA3.1-MALAT1-wt and corresponding control (pcDNA3.1) cells were selected by the culture medium containing 0.4 mg/ml Geneticin (G418, Invitrogen). After 6 weeks, G418-resistant cell clones were established to construct MALAT1 stable over-expression cell lines.

Wang Y., Zhang Y., Yang T., Zhao W., Wang N., Li P., Zeng X, & Zhang W. (2017). Long non-coding RNA MALAT1 for promoting metastasis and proliferation by acting as a ceRNA of miR-144-3p in osteosarcoma cells. Oncotarget, 8(35), 59417-59434.

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Transfection of Melanoma Cell Lines

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Melanoma cell lines A375, A2058, SK-MEL-5, SK-MEL-28 and human normal melanoma cell lines (MC) were purchased from the cell bank of the American Committee for Type Culture Collection. The cell culture medium consists of 10% fetal bovine serum solution (Gibco), 1% penicillin/streptomycin solution (100x, Solarbio) and DMEM basal medium (Hyclone). Cells were cultured in animal cell incubator (Binder, Germany) at 37°C and 5%CO₂ until they were in a good growth condition. Before transfection, the medium was replaced with fetal bovine serum-free medium. During transfection, 1×10⁵ cells per well were inoculated into 6-well plates. The miR-23a mimics, miR-23a inhibitor, MALAT1 siRNA and corresponding negative control vectors were all designed and synthesized by Shanghai Sangon Biotech Co., Ltd. Cell lines were transfected with Lipofectamine 2000 transfection kit (Invitrogen, USA). The procedures referred to the kit instructions. Eight hours after transfection, fresh culture medium was changed to avoid poisoning cells.

Wang P., Hu L., Fu G., Lu J., Zheng Y., Li Y, & Jia L. (2020). LncRNA MALAT1 Promotes the Proliferation, Migration, and Invasion of Melanoma Cells by Downregulating miR-23a. Cancer Management and Research, 12, 6553-6562.

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Modulating miR-2355-3p, IL6ST, and MALAT1 in Cellular Pathways

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miR-2355-3p mimic (100 nM), NC mimic (a negative control for miR-2355-3p mimic), miR-2355-3p inhibitor (100 nM), NC inhibitor (a negative control for miR-2355-3p inhibitor), IL6ST siRNA (100 nM), MALAT1 siRNA (100 nM), and NC siRNA or scramble (a negative control for IL6ST siRNA or MALAT1 siRNA) were synthesized by Invitrogen (Carlsbad, CA, United States). MALAT1 overexpression plasmid (pcDNA-lncRNA MALAT1) was synthesized by Thermo Fisher Scientific (Waltham, MA, United States), and pcDNA3.1 (Vector, a negative control for pcDNA-lncRNA MALAT1) was purchased from Invitrogen (Carlsbad, CA, United States). The transfection and co-transfection of these oligonucleotides and overexpression vectors were carried out with Lipofectamine Plus reagent (Invitrogen, Carlsbad, CA, United States) according to the manufacturer’s instructions.

Huang H., Zhang G, & Ge Z. (2021). lncRNA MALAT1 Promotes Renal Fibrosis in Diabetic Nephropathy by Targeting the miR-2355-3p/IL6ST Axis. Frontiers in Pharmacology, 12, 647650.

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Regulating MALAT1 and miR-23a-3p in HK-2 cells

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MALAT1 siRNA, MALAT1 siRNA NC, MALAT1 over-expression plasmid, miR-23a-3p mimic, miR-23a-3p inhibitor and the negative control were purchased from GenePharma (Shanghai, China). HK-2 cells were transfected by MALAT1 siRNA with or without miR-23a-3p inhibitor using Lipofectamine 3000 (Invitrogen) according to the protocol of the manufacturer.

Li J., Лю Б., & Li C. (2020). Long noncoding RNA MALAT1 can regulate proliferation and apoptosis of LPS-treated HK-2 cells via targeting miR-23a-3p through regulating ERK signaling. Archives of Medical Science.

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Targeted Knockdown of MALAT1 by siRNA in Cultured Cells

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Cells were plated in six-well plates at 2.0 × 10⁵ cells per well for 24 h with the serum-free medium before transfection. MALAT1 siRNA (Cat#: 4390771) and control negative siRNA (Mock, Cat#: 4390843) were obtained from Thermo Fisher Scientific. Lipofectamine™ RNAiMAX transfection reagent (Cat#: 13778075, Thermo Fisher Scientific) was used for transfection following the manufacturer’s instructions. Gene expression after siRNA knockdown was determined by PCR or quantitative reverse transcription-PCR (RT-Q-PCR) after 72 h of transfection.

Wu Y., Sarkissyan M., Ogah O., Kim J, & Vadgama J.V. (2020). Expression of MALAT1 Promotes Trastuzumab Resistance in HER2 Overexpressing Breast Cancers. Cancers, 12(7), 1918.

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Regulation of HUVEC Proliferation by miR-320a

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MALAT1 siRNAs were specifically designed and synthesized by Shanghai GenePharma Co, Ltd. (Shanghai, China) by targeting MALAT1 sequences (MALAT1-1, 5’-CACAGGGAAAGCGAGTGGTTGGTAA-3’ and 5’-TTACCAACCACTCGCTTTCCCTGTG-3’; and MALAT1-2, 5’-GAGGUGUAAAGGGAUUUAUTT-3’ and 5’-AUAAAUCCCUUUACACCUCTT-3’). A negative control siRNA was also synthesized with targeting sequences of 5’-GGCCUAAAGUAGUAGCUAUTT-3’ and 5’-AUAGCUACUACUUUAGGCCTT-3’.
To assess the effects of miR-320a on regulation of HUVEC proliferation, we had a miR-320a mimic, a miR-320a inhibitor, and control RNA chemically synthesized and purified using high-performance liquid chromatography (GenePharma). The concentration of these synthesized siRNAs (mimic, inhibitor, and negative control) were all 20 μM and the working concentration was 20 nM diluted at 1:1000. Plasmids containing MALAT1 siRNAs were transfected into cells for 48 h using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions and then the cells were used for our experiments.

Sun J.Y., Zhao Z.W., Li W.M., Yang G., Jing P.Y., Li P., Dang H.Z., Chen Z., Zhou Y.A, & Li X.F. (2017). Knockdown of MALAT1 expression inhibits HUVEC proliferation by upregulation of miR-320a and downregulation of FOXM1 expression. Oncotarget, 8(37), 61499-61509.

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MALAT1 siRNA Silencing in A549 Cells

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A total of 2 MALAT1 siRNAs (siRNA1 and siRNA2) and control siRNA were purchased from Invitrogen; Thermo Fisher Scientific, Inc. The siRNA sequences were as follows: MALAT1 siRNA1, 5′-GGGCUUCUCUUAACAUUUAtt-3′; MALAT1 siRNA2, 5′-GGGCAAAUAUUGGCAAUUAtt-3′. For the siRNA transfection, 2×10⁶ A549 or A549rCDDP cells were seeded on 6-well plates. The following day, 50 pM siRNA were mixed with Lipofectamine^® RNAiMAX (Invitrogen; Thermo Fisher Scientific, Inc.) in Opti-MEM (Thermo Fisher Scientific, Inc.) and incubated at 37°C for 15 min. Subsequently, the siRNA1 or siRNA2 mixture was added into the culture medium of cells and cultured at 37°C for 24 h prior to cisplatin or vehicle treatment.

Cui Y., Li G., Zhang X., Dai F, & Zhang R. (2018). Increased MALAT1 expression contributes to cisplatin resistance in non-small cell lung cancer. Oncology Letters, 16(4), 4821-4828.

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