Synthetic RNA oligonucleotides targeting H19 were obtained from RiboBio (Guangzhou, China). The siRNA target sequence was GCGGGTCTGTTTCTTTACT. pcDNA3.1-H19 was purchased from GenePharma (Shanghai, China). miR675-3p-mimics and miR675-3p-inhibitor were obtained from RiboBio (Guangzhou, China). HEK293T cells were transfected with si-H19, pcDNA3.1-H19, miR675-3p-mimics, and miR675-3p-inhibitor for 48 h, respectively. Control cells were transfected with nonspecific, scrambled siRNA.
Pcdna3.1 h19
Manufactured by GenePharma
Sourced in China
PcDNA3.1-H19 is a plasmid vector designed for the expression of the H19 gene in mammalian cells. The vector contains the necessary elements for gene expression, including a promoter and a polyadenylation signal. PcDNA3.1-H19 can be used for various research applications that involve the study of the H19 gene and its expression in cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Mir 140 5p mimic, by GenePharma (3 mentions)
Lenti shnc, by GenePharma (3 mentions)
Shh19 1, by GenePharma (3 mentions)
Shh19 2, by GenePharma (3 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (3 mentions)
Synthetic rna oligonucleotides targeting h19, by RiboBio (2 mentions)
Pcdna3.1 empty vector, by GenePharma (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Mir675 3p mimics, by RiboBio (1 mentions)
Si nc, by GenePharma (1 mentions)
Pcdna3, by GenePharma (1 mentions)
Fopflash, by Merck Group (1 mentions)
Prl sv40 plasmid, by Promega (1 mentions)
Synergy h2 microplate reader, by Agilent Technologies (1 mentions)
Dual luciferase reporter assay system, by Promega (1 mentions)
α mem, by GenePharma (1 mentions)
Scramble control nc, by GenePharma (1 mentions)
Si h19, by RiboBio (1 mentions)
9 protocols using pcdna3.1 h19
1
Investigating the Role of H19 and miR675-3p in Cell Lines
2
Manipulating H19 and miR-29a-3p in Osteoclasts
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pcDNA3.1-H19, pcDNA3.1, H19 targeting siRNA (si-H19) and its negative control (si-NC) were purchased from Genepharma Inc. (Shanghai, China). Osteoclasts transfection with pcDNAs or siRNAs were carried out by Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacture protocol. The miR-29a-3p inhibitor and negative control was bought from Genepharma.
3
Dual Luciferase Assay for H19 lncRNA
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The TOP Flash and FOP Flash plasmids used in this study were purchased from Millipore (USA) and the pRLSV40 plasmid (Promega, USA) containing the Renilla reniformis luciferase gene (RLuc) under the control of the SV40 virus promoter (kindly provided by Professor Bu in the Peking University First Hospital Central Laboratory) was used as control. The expressing plasmids pcDNA 3.1(+)-H19 and pcDNA 3.1(+) empty vector were purchased from Genepharma (China). Cells were transfected with 500 ng H19 or empty vector, 500 ng TOP Flash or FOP Flash vector, and 12 ng RLuc using Lipofectamine 3000.12 h after transfection, cells were incubated with indicated doses of 1,25(OH)2D3 or equal quantities of ethanol for 48 and the analysis of Luc and RLuc activities was performed using the Dual Luciferase Reporter Assay System (Promega, USA) and the Luc and Rluc activities were measured with Synergy H2 microplate reader (Bio Tek Instruments, USA). The Luc activity was then normalized to RLuc activity.
4
Stable H19 Overexpression in Colon Cancer Cells
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To establish cells stably overexpressing H19, HT-29 and DLD-1 cells were transfected with pcDNA 3.1 (+)-H19 (GenePharma, China) or pcDNA 3.1(+) empty vector (GenePharma, China) as control using Lipofectamine 3000 and cells were allowed to recover for 48 h. Cell were then incubated with medium containing G418 (Sigma Aldrich, USA) (1.8 mg/ml for HT-29 and 1 mg/ml for DLD-1) for at least 4 weeks. The expression level of H19 of clones obtained were investigated and the clones with highest expression level of H19 were used in the studies both in vitro and in vivo.
5
Lentiviral-mediated Modulation of H19 and miR-140-5p in hDPSCs
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Recombinant lentiviruses targeting H19 (shH19-1 and shH19-2) and Lenti-shNC were purchased from GenePharma Company (Shanghai, China). hDPSCs were transfected by lentiviruses exposure in 1 mL α-MEM supplemented with 10% FBS and 5 μg/mL polybrene for 24 h. H19 overexpression plasmid pcDNA3.1-H19, miR-140-5p mimics, and scramble control (NC) were chemically synthesized by GenePharma. When hDPSCs were 70–80% confluent, pcDNA-H19 and miRNA mimic transfection was performed using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer’s instructions. qRT-PCR analysis was used to detect H19 and miR-140-5p expression levels to validate the transfection efficiencies. The cells were cultured in mineralizing medium for odontoblastic differentiation 48 h after transfection.
6
Plasmid and RNA Modulation of H19
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pcDNA3.1-H19, a plasmid for H19 overexpression, was sourced from the GenePharma (Shanghai, China). si-H19, a small interfering RNA that reduces H19 expression, was obtained from RiboBio. miR-675-5p inhibitors and mimics, which can reduce or overexpress miR-675-5p, were obtained from RiboBio.
7
Modulating H19 and miR675-3p in Liver Cancer Cells
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Synthetic RNA oligonucleotides targeting H19 was obtained from RiboBio (Guangzhou). The siRNA target sequence was GCGGGTCTGTTTCTTTACT. pcDNA3.1‐H19 was procured from GenePharma (Shanghai, China). miR675‐3p mimics and inhibitor were obtained from RiboBio (Guangzhou).
The CRISPR/Cas9 plasmids were obtained from Addgene (px458). Protocols for sgRNA design and the procedures required for the in vitro transcription have been described previously.20 The sgRNA‐oligo sequences are listed in Table S1 .
MHCC97H cells were transfected with si‐H19, pcDNA3.1‐H19, miR675‐3p‐mimics, miR675‐3p‐inhibitor, H19‐KO for 48 hours, respectively. Control cells were transfected with nonspecific or scrambled siRNA.
The CRISPR/Cas9 plasmids were obtained from Addgene (px458). Protocols for sgRNA design and the procedures required for the in vitro transcription have been described previously.
MHCC97H cells were transfected with si‐H19, pcDNA3.1‐H19, miR675‐3p‐mimics, miR675‐3p‐inhibitor, H19‐KO for 48 hours, respectively. Control cells were transfected with nonspecific or scrambled siRNA.
8
Lentiviral-mediated H19 Regulation in hDPSCs
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Recombinant lentiviruses targeting H19 (shH19-1 and shH19-2) and Lenti-shNC were purchased from GenePharma Company (Shanghai, China). hDPSCs were transfected by lentiviruses exposure in 1 mL α-MEM supplemented with 10% FBS and 5μg /mL polybrene for 24 h. H19 overexpression plasmid pcDNA3.1-H19, miR-140-5p mimics and and scramble control (NC) were chemically synthesized by GenePharma. When hDPSCs were 70%-80% con uent, pcDNA-H19 and miRNA mimic transfection was performed using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer's instructions. qRT-PCR analysis was used to detect H19 and miR-140-5p expression levels to validate the transfection e ciencies. The cells were cultured in mineralizing medium for odontoblastic differentiation 48 h after transfection.
9
Modulating H19 and miR-140-5p in hDPSCs
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Recombinant lentiviruses targeting H19 (shH19-1 and shH19-2) and Lenti-shNC were purchased from GenePharma Company (Shanghai, China). hDPSCs were transfected by lentiviruses exposure in 1 mL α-MEM supplemented with 10% FBS and 5 µg /mL polybrene for 24 h. H19 overexpression plasmid pcDNA3.1-H19, miR-140-5p mimics and and scramble control (NC) were chemically synthesized by GenePharma. When hDPSCs were 70%-80% con uent, pcDNA-H19 and miRNA mimic transfection was performed using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer's instructions. qRT-PCR analysis was used to detect H19 and miR-140-5p expression levels to validate the transfection e ciencies. The cells were cultured in mineralizing medium for odontoblastic differentiation 48 h after transfection.
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