To overexpress or suppress miR-126-3p in hCECs, RNA oligonucleotides for miR-126 agomir and antagomir were synthesized by GenePharm, Shanghai, People’s Republic of China, and a scrambled oligonucleotide served as the control. When the cells reached a confluence of 30%–40% in six-well plates, the normal hCECs were transfected with a final concentration of 5 nM of RNA oligonucleotides using a transfection agent purchased from GenePharm. After 6 h, the cells were washed three times with PBS and were induced in total EGM-2 medium for another 4 days. The hCECs from the NC group, DM group, and DM+ICA II group were induced in the corresponding media as previously described, and ICA II, at a final concentration of 10 μM, was used in this assay.
Rna oligonucleotides
Manufactured by GenePharma
Sourced in China
RNA oligonucleotides are short, synthetic RNA molecules used in various research and therapeutic applications. They serve as essential tools for studying gene expression, regulation, and function. RNA oligonucleotides can be designed to target specific RNA sequences, enabling researchers to investigate and manipulate cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (2 mentions)
Interferin reagent, by Polyplus Transfection (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Mirna inhibitor, by GenePharma (1 mentions)
Mirna mimic, by GenePharma (1 mentions)
Cmv promoter, by GenePharma (1 mentions)
Sirnas, by GenePharma (1 mentions)
Rna oligonucleotide, by Horizon Discovery (1 mentions)
Dharmafect sirna transfection reagent, by Horizon Discovery (1 mentions)
Rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions)
Block it rnai designer, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (1 mentions)
Anti mir nc, by GenePharma (1 mentions)
Opti mem 1 reduced serum media, by Thermo Fisher Scientific (1 mentions)
17 protocols using rna oligonucleotides
1
Modulating miR-126-3p in hCECs
2
Comprehensive Molecular Toolkit for lnc-UCID Research
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All RNA oligonucleotides were purchased from GenePharma (Shanghai, China). The small interfering RNAs (siRNAs) targeting the human lnc‐UCID (Ensembl transcript ID: ENST00000497831.1), DHX9 (GeneBank accession No. NM_001357.4), and CDK6 (NM_001259) transcripts were designated as siUCID, siDHX9 and siCDK6, respectively. The negative control (NC) RNA duplex for both miR‐148a and siRNAs was nonhomologous to any human genome sequence. The miR‐148a inhibitor (anti‐miR‐148a) with a sequence complementary to the mature miR‐148a and its negative control (anti‐NC) consisted of 2′‐O‐methyl‐modified RNA oligonucleotides.
The expression vectors pc3‐puro‐UCID, pc3‐puro‐UCID‐Δcore, pc3‐gab‐UCID, pc3‐gab‐CDK6, pCDH‐Flag‐DHX9, and luciferase reporter vectors psi‐CDK6‐DHX9‐binding element (DBE) 1, psi‐CDK6‐DBE2, psi‐UCID‐WT, and psi‐UCID‐MUT were generated as described in the Supporting Information.
All oligonucleotide sequences are listed in Supporting TableS2 .
The expression vectors pc3‐puro‐UCID, pc3‐puro‐UCID‐Δcore, pc3‐gab‐UCID, pc3‐gab‐CDK6, pCDH‐Flag‐DHX9, and luciferase reporter vectors psi‐CDK6‐DHX9‐binding element (DBE) 1, psi‐CDK6‐DBE2, psi‐UCID‐WT, and psi‐UCID‐MUT were generated as described in the Supporting Information.
All oligonucleotide sequences are listed in Supporting Table
3
Transfection of Oligonucleotides and Plasmids in C2C12 Cells
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All the RNA oligonucleotides in this study (miRNA mimics, miRNA inhibitor, and siRNAs) were obtained from GenePharma (Suzhou, China). RNA oligonucleotides’ sequences are listed in Additional file 2 : Table S2. The full length of CamkIIδ coding sequence was cloned into pcDNA3.1 and driven to be expressed by CMV promoter (GenePharma, Suzhou, China). For proliferation, cells were seeded into 96 or 24-well plates before transfection and transfected with Lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instruction when reached approximately 30% confluence. For myogenic differentiation assay, C2C12 cells were seeded in 12-well plates and transfected when reached 70% confluence. RNA oligonucleotides applied in cell transfection were described in Additional file 2 : Table S2.
4
Silencing STAT3 and β‐Arrestin1 in Cells
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The RNA oligonucleotides that specifically targeted rat STAT3, β‐arrestin1 were purchased from GenePharma (Shanghai, China). The RNA oligonucleotides were transfected with the Dharma‐FECT siRNA transfection reagent (Dharmacon). After siRNA transfection for 24 or 48 h, cells were harvested and analyzed by western blotting and qRT‐PCR.
5
Investigating miR-33-5p's Regulatory Role
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In this experiment, total sequences of DNA primers were adopted, which are presented in Table 1 Table 2
6
Endogenous siRNA Duplex Synthesis
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RNA oligonucleotides (the guide and passenger strands) of each siRNA were chemically synthesized (GenePharma) and annealed to form endogenous siRNA duplexes. The siRNA sequences are shown in Supplementary Table S1 .
7
RNA Oligonucleotides for MiRNA and siRNA Transfection
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RNA oligonucleotides were chemically synthesized and purified by Genepharma Co. Ltd., (Shanghai, China). Sequence of human miR-30b mimics was 5′- UGU AAA CAUC CUA CAC UCA GCU -3′ and human miR-30c mimics was 5′- UGU AAA CAU CCU ACA CUC UCA GC -3′. Negative control oligonucleotides for miRNA mimics was 5′-CAG UAC UUU UGU GUA GUA CAA-3′. The sequences of Rab18 siRNA was: 5′- GAA ACA UAC UGU ACA AGA ATT -3′ (sense) and 5′-UUC UUG UAC AGU AUG UUU CTT-3′ (antisense), Control siRNA was: 5′-UUC UCC GAA CGU GUC ACG UTT-3′ (sense), 5′-ACG UGA CAC GUU CGU AGA ATT-3′ (antisense). The transfections were performed with INTERFERin reagent (Polyplus-transfection). The final concentration of miRNA was 50 nM. The final concentration of siRNA was 20 nM.
8
Transfection of miRNA Inhibitors and Mimics
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RNA oligonucleotides were purchased from Genepharma Co., Ltd (Shanghai, China). The human miR-17 inhibitors sequence was 5′-CUACCUGCACUGUAAGCACUUUG-3′. The NC oligonucleotides sequence for miR-17 was 5′-CAGUACUUUUGUGUAGUACAA-3′. miR-17 mimics sequence was 5′-CAAAGUGCUUACAGUGCAGGUAG-3′, The negative control (NC) sequence was 5′-UUCUCCGAACGUGUCACGUTT-3′. INTERFERin reagent (89129-130, Polyplus Transfection) was used to conduct the transfection. The final concentration of miRNA inhibitors or miR-17 mimics or NC was 50 nM, and the final concentration of siRNAs or NC was 20 nM.
9
siRNA Knockdown of Cell Trafficking Proteins
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RNA oligonucleotides used for siRNA were purchased from GenePharma Co., Ltd. The sequences were as follows: NC, 5′-UUCUCCGAACGUGUCACGUTT-3′, SNX27, 5′-CCAGGUAAUUGCAUUUGAATT-3′, VPS35, 5′-AAAUACCAGUUGACACUUATT-3′, KIAA1033-1, 5′-GCCCUCUAUAUCAGUAACATT-3′, KIAA1033-2, 5′-GAGCUGUCUUCCCAAUUUATT-3′, KIAA1033-3, 5′-GGCCAAAGAAUAUACAUCUTT-3′.
Cells grown in 6-well plates or glass-bottom dishes were transfected with 100 pmol RNA oligonucleotides twice at an interval of 24 h using Lipofectamine 2000 (Invitrogen). Cells were harvested for further experiments 72 h after the first transfection.
Cells grown in 6-well plates or glass-bottom dishes were transfected with 100 pmol RNA oligonucleotides twice at an interval of 24 h using Lipofectamine 2000 (Invitrogen). Cells were harvested for further experiments 72 h after the first transfection.
10
RNA Interference Assay Protocol
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RNA oligonucleotides (GenePharma, Shanghai, China) (Table 1
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