Pcdna3.1 plasmid

Manufactured by GenePharma

Sourced in China

PcDNA3.1 plasmid is a circular DNA vector commonly used in molecular biology experiments. It contains elements necessary for replication and selection in both bacterial and mammalian cells, including an ampicillin resistance gene and a cytomegalovirus (CMV) promoter for high-level transgene expression.

Automatically generated - may contain errors

Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (17 mentions) Lipofectamine 3000 reagent, by Thermo Fisher Scientific (6 mentions) Lipofectamine 2000, by Highfine Biotech (6 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (4 mentions) Si nc, by RiboBio (3 mentions) Sh nc, by GenePharma (2 mentions) Anti mir nc, by RiboBio (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Lipofectamine reagent, by Thermo Fisher Scientific (2 mentions) Ribofect cp reagent, by RiboBio (2 mentions) Sh usp7, by GenePharma (1 mentions) Mir nc inhibitor, by RiboBio (1 mentions) App ps1 double transgenic mice, by Jackson ImmunoResearch (1 mentions) Mir 27a mimic, by GenePharma (1 mentions) Mir 27a inhibitor, by GenePharma (1 mentions) Mir 637 mimic and inhibitor, by GenePharma (1 mentions) Scrambled control sirna, by RiboBio (1 mentions) Puromycin, by Merck Group (1 mentions) Sh linc00689, by GenePharma (1 mentions)

35 protocols using pcdna3.1 plasmid

Modulating HOTAIR and PKR Expression

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Short-hairpin RNA directed against HOTAIR was ligated into the pcDNA3.1 plasmid
(GenePharma, China) and was referred to as sh-HOTAIR. The sh-HOTAIR sequences
were as follows: sense,
5′-GATCCGCCACATGAACGCCCAGAGATTTTCAAGAGAAATCTCTGGGCGTTCATGTGGTTTTTTG-3′, and
anti-sense,
5′-AATTCAAAAAACCACATGAACGCCCAGAGATTTCTCTTGAAAATCTCTGGGCGTTCATGTGGCG-3′. The
plasmid carrying non-targeting sequence was used as NC of sh-HOTAIR and was
referred to as sh-NC. Plasmids overexpressing HOTAIR were ligated into the
pcDNA3.1 and referred to as pc-HOTAIR. The full-length PKR sequences were
constructed into pcDNA3.1 plasmids (GenePharma) to analyze the functions of PKR,
and were referred to as pc-PKR. PKR siRNA was obtained from Bioneer (Daejeon
Korea) with a sequence of 5′-CGUUGCUUAUGAAUGGUCU-3′. Lipofectamine 3000 reagent
(Life Technologies Corporation, USA) was used for cell transfection according to
the manufacturer's instructions. The stably transfected cells were selected by
the culture medium containing 0.5 mg/mL G418 (Sigma-Aldrich, USA). After
approximately 4 weeks, G418-resistant cell clones were established.

Liu G, & Zhang W. (2018). Long non-coding RNA HOTAIR promotes UVB-induced apoptosis and inflammatory injury by up-regulation of PKR in keratinocytes. Brazilian Journal of Medical and Biological Research, 51(8), e6896.

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Modulating MTDH and USP7 Expression

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The small interference RNA (siRNA) targeting MTDH or USP7 (si-MTDH or si-USP7) or the short hairpin RNA (shRNA) targeting USP7 (sh-USP7) were designed by GenePharma with nontargeted siRNAs or shRNAs as the negative control (si-NC or sh-NC). MTDH-overexpression plasmids (MTDH) were established by cloning full-length of MTDH into pcDNA3.1 plasmids (GenePharma), and the empty pcDNA3.1 plasmids were used as the negative control (pcDNA). The transient transfection was performed using the Lipofectamine 3000 (Invitrogen).

Wang N., Xu J., Wang Y., Zhang X, & Zhang H. (2024). USP7 promotes cervical cancer progression by stabilizing MTDH expression through deubiquitination. Journal of Cancer Research and Clinical Oncology, 150(4), 196.

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Regulation of RAB10 expression

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MiR-557 mimic, miR-557 inhibitor, miR-mimic negative control (NC), miR-inhibitor-NC, and the siRNA targeting RAB10 and NC (siRAB10, siNC) were acquired from RiboBio (Guangzhou, China). For RAB10 up-expression, cDNA of RAB10 was amplified by PCR and then inserted into the pcDNA3.1(+) plasmids (GenePharma, Shanghai, China). All of them were infected into cells with Lipofectamine 2000 (Invitrogen, USA). The infection rate was validated by RT-qPCR. The sequences of cDNA clone and siRNA are displayed in Supplementary Table 1.

Cheng X., Wu C., Xu H., Zou R., Li T, & Ye S. (2024). miR-557 inhibits hepatocellular carcinoma progression through Wnt/β-catenin signaling pathway by targeting RAB10. Aging (Albany NY), 16(4), 3716-3733.

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Alzheimer's Disease Model Mice Studied

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The APP/PS1 double transgenic mice were purchase from Jackson Laboratory and selected as the AD models. All these mice were allowed access to free food and water to get used to the new situation in an animal room meeting SPF standards. The growth temperature was 24°C with a relative humidity of 55% ∼ 65% and daily light or night for 12 h. All the experimental schemes used in this study have been approved by the Committee on Ethics of Experimental Animals. All animals were divided into controls, voluntary exercise (VE) cohort, exercise and lncRNA negative control (NC) group, and exercise and p-SNHG14 group. A total of 10 mice were included in each group. Mice in the control group were housed in the cages with fixed wheels and mice in other groups were housed in the cages with running wheels. The sequences of SNHG14 or its NC were cloned into pcDNA3.1 plasmids (GenePharma, Shanghai, China) and then recombinant plasmids (p-NC or p-SNHG14) were injected into the bilateral hippocampus of APP/PS1 mice. After 4-week of training, half the mice of each group were selected randomly to be euthanized, the rest rice received the following experiments. The graphical timeline of experimental mice was shown in Figure 1A.

He Y, & Qiang Y. (2021). Mechanism of Autonomic Exercise Improving Cognitive Function of Alzheimer’s Disease by Regulating lncRNA SNHG14. American Journal of Alzheimer's Disease and Other Dementias, 36, 15333175211027681.

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Modulating circ_0043947 and miR-384

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Small interfering RNAs (siRNAs) or short hairpin RNA (shRNA) targeting circ_0043947 (si-circ_0043947 and sh-circ_0043947), miR-384 mimics and inhibitors (miR-384 and anti-miR-384) and the corresponding controls were provided by RiboBio (Guangzhou, China). The CREB1 overexpression plasmid (pcDNA-CREB1) was constructed with pcDNA3.1 plasmids (GenePharma, Shanghai, China).

Zhang C., Zhang F., Li Y., Yang P., Liu Y, & Yang W. (2024). The Circular RNA Circ_0043947 Promoted Gastric Cancer Progression by Sponging miR-384 to Regulate CREB1 Expression. Gut and liver.

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Genetic Manipulation of miR-27a and NELL-1

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miR-27a mimic, miR-27a inhibitor, and their negative control (NC, scrambled) were synthesized by GenePharma Co. (China). For overexpressing NELL-1, full-length wild type NELL-1 sequences were constructed into pcDNA3.1 plasmids (GenePharma). Empty pcDNA3.1 plasmid was used as a blank control. For NELL-1 silencing, NELL-1-targeted specific siRNAs (GenePharma) were transfected into cells. Non-targeting sequences were used as a blank control. Cell transfections were conducted using Lipofectamine 3000 reagent (Invitrogen, USA) following the manufacturer's protocol. At 48 h of transfection, 0.5 mg/mL G418 (Sigma-Aldrich) was added to the culture medium for selection of stably transfected cells. After approximately 4 weeks, G418-resistant cell clones were established and were collected for use in the following experiments.

Chen H., Zhang Z., Zhang L., Wang J., Zhang M, & Zhu B. (2018). miR-27a protects human mitral valve interstitial cell from TNF-α-induced inflammatory injury via up-regulation of NELL-1. Brazilian Journal of Medical and Biological Research, 51(6), e6997.

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Overexpression and Knockdown of circUSP36 and WNT4

Cited 1 time

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circUSP36-overexpression vectors were constructed by cloning the transcript sequence of circUSP36 into pcDNA3.1 plasmids obtained from Genepharma (Shanghai, China). Small interfering RNAs (siRNA) against WNT4 or circUSP36 for knockdown of WNT4 or circUSP36 (named as si-WNT4 and si-circUSP36, respectively) and their negative control the scramble siRNA (named as si-NC) were designed and supplied by RiboBio Co., Ltd. (Guangzhou, China). To regulate miR-637 expression, oligonucleotides including miR-637 mimic and inhibitor, as well as their corresponding mimic NC and inhibitor NC, were acquired from Genepharma. Cell transfection was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the instructions [25 (link)].

Huang J.G., Tang X., Wang J.J., Liu J., Chen P, & Sun Y. (2021). A circular RNA, circUSP36, accelerates endothelial cell dysfunction in atherosclerosis by adsorbing miR-637 to enhance WNT4 expression. Bioengineered, 12(1), 6759-6770.

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HepG2 Cell Line Silencing HBx and XB130

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Human liver cancer cell line HepG2 and normal hepatocyte MIHA were incubated in Roswell Park Memorial Institute-1640 medium at 37°C with 5% CO₂. The cell suspension of HepG2 cells was cultured in a 37°C incubator with 5% CO₂ for 24 h, after which cells were seeded onto culture dishes for overnight culture. The plasmids containing small interfering RNA targeting HBx (si-HBx) and XB130 (si-XB130) fragments and empty plasmids were transfected into HepG2 cells using Lipofectamine 2000 (Thermo Fisher Scientific) when cell confluence reached 60%. At 4 h posttransfection, the medium was renewed, and cells were further incubated for 48 h for subsequent experiments. The pcDNA3.1 plasmids (GenePharma, Shanghai, China) were used, while the sequence fragments were synthesized by Sangon Biotechnology (Shanghai, China).

Huang C., Liu W., Zhao X., Zhao L, & Wang F. (2021). Downregulation of HBx Restrains Proliferation, Migration, and Invasion of HepG2 Cells. Analytical Cellular Pathology (Amsterdam), 2021, 6615979.

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Cloning and Knockdown of Regulatory Genes

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Full-length human lincROR, MYH9, HIF1α, and hnRNPA1 were amplified and cloned into the pcDNA3.1 plasmids (GenePharma). siRNA targeting lincROR, MYH9, HIF1α, hnRNPA1, and scrambled control siRNA were synthesized by RiboBio. The cells were transfected using Lipofectamine 2000 (Invitrogen). The small hairpin RNA (shRNA) sequence of lincROR was cloned into GV112 plasmid (GenePharma), and lentiviral particles were harvested and transduced in the presence of polybrene. The knockdown (KD) stable cell lines were obtained by selection of transduced cells with 2 μg/mL of puromycin (Sigma-Aldrich).

Jiang X., Xu Y., Liu R, & Guo S. (2023). Exosomal lincROR Promotes Docetaxel Resistance in Prostate Cancer through a β-catenin/HIF1α Positive Feedback Loop. Molecular cancer research : MCR, 21(5).

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Gastric Cancer Cell Culture Transfection

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Negative control (NC), miR-526b-3p mimics, miR-526b-3p inhibitor, sh-NC, sh-LINC00689, si-ADAM9, pcDNA3.1 plasmid and pcDNA3.1-LINC00689 were all obtained from Genepharma (Shanghai, China). The cell suspension was prepared by 0.25% pancreatin digested AGS or MGC-803 cells and complete medium. Then, the cells were incubated in six-well plates (1×10⁶ cells/well) for 18–24 h. Three hours before transfection, GC cells at 80–90% confluence were incubated with fresh medium without serum or antibiotics. Transfection was performed using Lipofectamine 2000 (~ 0.6 μg Lipofectamine reagent/1 μg DNA, Life Technologies).

Yin G., Tian P., BuHe A., Yan W., Li T, & Sun Z. (2020). LncRNA LINC00689 Promotes the Progression of Gastric Cancer Through Upregulation of ADAM9 by Sponging miR-526b-3p. Cancer Management and Research, 12, 4227-4239.

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