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PureFection Transfection Reagent

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[#LV750A-1] PureFection Transfection Reagent

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(1) MiR-136 triggers apoptosis in human gastric cancer cells by targeting AEG-1 and BCL2.[TOP]

Pubmed ID :30468468
Publication Date : //
Gastric cancer is the second most prevalent cancer across the globe and accounts for about 10% of new cancer cases. It is one among the leading causes of cancer-related deaths around the world. Recently, microRNAs have been identified as important therapeutic targets for the treatment of several cancers owing to their potential to target multiple genes and hinder several biological processes such as proliferation and apoptosis. In the current study, we investigated the potential of miR-136 as therapeutic target for gastric cancer.

Authors : Yu L, Zhou G-Q, Li D-C,



(2) Application of ion pair chromatography coupled with mass spectrometry to assess antisense oligonucleotides concentrations in living cells.[TOP]

Pubmed ID :30462105
Publication Date : //
Antisense oligonucleotides (ASOs) are synthetic bioactive compounds used as therapeutic agents in clinical trials. They act by binding to complementary sequences of the targeted nucleic acids in cells. Assessing the efficiency of ASO delivery to cells or tissues and the stability of these compounds in different biological systems is important. To answer these questions, we developed a new, quick and reliable method to determine the concentrations of different types of ASOs in treated cells. Ultra-high performance liquid chromatography coupled with mass spectrometry was used for the first time for the separation and determination of the studied compounds in total RNA extracts. To develop a method with the highest possible sensitivity, a central composite design was used to comprehensively optimize the MS parameters. Moreover, the effects of the type and concentration of the ion pair reagent on sensitivity were also examined. Finally, a mobile phase containing methanol, hexafluoroisopropanol and N,N-dimethylbutylamine was selected. The optimized method allowed good linearity, accuracy, precision and sensitivity of ASO detection. Next, these compounds were delivered into cells via transfection at a concentration of 25 nM or 125 nM in 1 mL of cell culture medium. After 48 hours, total RNA was isolated from the treated cells and analyzed with the use of the newly developed method. For the cells treated with a higher concentration of ASO composed of phosphorothioate 2'-O-methyl RNA units, the concentration in solution was 0.96 ± 0.06 μM, while in the case of shorter ASO composed of locked nucleic acid units, it was 0.72 ± 0.06 μM in the total RNA extract.

Authors : Studzińska Sylwia, Cywoniuk Piotr, Sobczak Krzysztof,



(3) Enhanced uptake of plasmid at boronic acid decorated linear polyethylenimines results in higher transfection efficiency.[TOP]

Pubmed ID :30458622
Publication Date : //
High molecular weight polyethylenimines (PEIs) are considered as gold standard for transfection studies; however, cytotoxicity associated with branched ones and lower charge density on linear PEIs as well as lower uptake of the resulting deoxyribonucleic acid (DNA) complexes have limited their applications in clinical studies. In order to address these concerns and improve the uptake efficiency of the DNA complexes of linear polyethylenimine (25 kDa), the polymer was grafted with variable amounts of butylboronic acid to obtain a small series of linear polyethylenimine-butylboronic acid polymers. These modified polymers were allowed to interact with plasmid DNA and the resulting complexes were characterized by physicochemical techniques. Dynamic light scattering data showed the formation of nanosized complexes with positive zeta potential values. Furthermore, when these complexes were evaluated , they not only showed enhanced cell viability but also exhibited higher transfection efficiency as compared to native linear and branched PEIs and a commercially available standard transfection reagent, Lipofectamine 2000.

Authors : Yadav Santosh, Kumar Pradeep,



(4) Dual carrier-cargo hydrophobization and charge ratio optimization improve the systemic circulation and safety of zwitterionic nano-polyplexes.[TOP]

Pubmed ID :30458360
Publication Date : //
While polymeric nano-formulations for RNAi therapeutics hold great promise for molecularly-targeted, personalized medicine, they possess significant systemic delivery challenges including rapid clearance from circulation and the potential for carrier-associated toxicity due to cationic polymer or lipid components. Herein, we evaluated the in vivo pharmacokinetic and safety impact of often-overlooked formulation parameters, including the ratio of carrier polymer to cargo siRNA and hydrophobic siRNA modification in combination with hydrophobic polymer components (dual hydrophobization). For these studies, we used nano-polyplexes (NPs) with well-shielded, zwitterionic coronas, resulting in various NP formulations of equivalent hydrodynamic size and neutral surface charge regardless of charge ratio. Doubling nano-polyplex charge ratio from 10 to 20 increased circulation half-life five-fold and pharmacokinetic area under the curve four-fold, but was also associated with increased liver enzymes, a marker of hepatic damage. Dual hydrophobization achieved by formulating NPs with palmitic acid-modified siRNA (siPA-NPs) both reduced the amount of carrier polymer required to achieve optimal pharmacokinetic profiles and abrogated liver toxicities. We also show that optimized zwitterionic siPA-NPs are well-tolerated upon long-term, repeated administration in mice and exhibit greater than two-fold increased uptake in orthotopic MDA-MB-231 xenografts compared to commercial transfection reagent, in vivo-jetPEI. These data suggest that charge ratio optimization has important in vivo implications and that dual hydrophobization strategies can be used to maximize both NP circulation time and safety.

Authors : Jackson Meredith A, Bedingfield Sean K, Yu Fang, Stokan Mitchell E, Miles Rachel E, Curvino Elizabeth J, Hoogenboezem Ella N, Bonami Rachel H, Patel Shrusti S, Kendall Peggy L, Giorgio Todd D, Duvall Craig L,



(5) The Effect of the MicroRNA-183 Family on Hair Cell-Specific Markers of Human Bone Marrow-Derived Mesenchymal Stem Cells.[TOP]

Pubmed ID :30380528
Publication Date : //
Hearing loss is considered the most common sensory disorder across the world. Nowadays, a cochlear implant can be an effective treatment for patients. Moreover, it is often believed that sensorineural hearing loss in humans is caused by loss or disruption of the function of hair cells in the cochlea. In this respect, mesenchymal cells can be a good candidate for cell-based therapeutic approaches. To this end, the potential of human bone marrow-derived mesenchymal stem cells to differentiate into hair cells with the help of transfection of microRNA in vitro was investigated. MicroRNA mimics (miRNA-96, 182, and 183) were transfected to human bone marrow-derived mesenchymal stem cells using Lipofec-tamine as a common transfection reagent following the manufacturer's instructions at 50 nM for microRNA mimics and 50 nM for the scramble. The changes in cell morphology were also observed under an inverted microscope. Then, the relative expression levels of SOX2, POU4F3, MYO7A, and calretinin were assayed using real-time polymerase chain reaction according to the ΔΔCt method. The ATOH1 level was similarly measured via real-time polymerase chain reaction and Western blotting. The results showed that increased expression of miRNA-182, but neither miRNA-96 nor miRNA-183, could lead to higher expression levels in some hair cell markers. The morphology of the cells also did not change in this respect, but the evaluation of gene expression at the levels of mRNA could promote the expression of the ATOH1, SOX2, and POU4F3 markers. Furthermore, miRNA-182 could enhance the expression of ATOH1 at the protein level. According to the results of this study, it was concluded that miRNA-182 could serve as a crucial function in hair cell differentiation by the upregulation of SOX2, POU4F3, and ATOH1 to promote a hair cell's fate.

Authors : Mahmoudian-Sani Mohammad-Reza, Jami Mohammad-Saeid, Mahdavinezhad Ali, Amini Razieh, Farnoosh Gholamreza, Saidijam Massoud,



(6) Establishment of a rapid and footprint-free protocol for differentiation of human embryonic stem cells into pancreatic endocrine cells with synthetic mRNAs encoding transcription factors.[TOP]

Pubmed ID :30359326
Publication Date : //
Transplantation of pancreatic β cells generated in vitro from pluripotent stem cells (hPSCs) such as embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) has been proposed as an alternative therapy for diabetes. Though many differentiation protocols have been developed for this purpose, lentivirus-mediated forced expression of transcription factors (TF)-PDX1 and NKX6.1-has been at the forefront for its relatively fast and straightforward approach. However, considering that such cells will be used for therapeutic purposes in the future, it is desirable to develop a procedure that does not leave any footprint on the genome, as any changes of DNAs could potentially be a source of unintended, concerning effects such as tumorigenicity. In this study, we attempted to establish a novel protocol for rapid and footprint-free hESC differentiation into a pancreatic endocrine lineage by using synthetic mRNAs (synRNAs) encoding PDX1 and NKX6.1. We also tested whether siPOU5F1, which reduces the expression of pluripotency gene POU5F1 (also known as OCT4), can enhance differentiation as reported previously for mesoderm and endoderm lineages.

Authors : Ida Hideomi, Akiyama Tomohiko, Ishiguro Keiichiro, Goparaju Sravan K, Nakatake Yuhki, Chikazawa-Nohtomi Nana, Sato Saeko, Kimura Hiromi, Yokoyama Yukihiro, Nagino Masato, Ko Minoru S H, Ko Shigeru B H,



(7) Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles.[TOP]

Pubmed ID :30323594
Publication Date : //
The delivery of transgenes into human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (hiPSC-CMs) represents an important tool in cardiac regeneration with potential for clinical applications. Gene transfection is more difficult, however, for hiPSCs and hiPSC-CMs than for somatic cells. Despite improvements in transfection and transduction, the efficiency, cytotoxicity, safety, and cost of these methods remain unsatisfactory. The objective of this study is to examine gene transfection in hiPSCs and hiPSC-CMs using magnetic nanoparticles (NPs).

Authors : Yamoah Megan A, Moshref Maryam, Sharma Janhavi, Chen Wei Chun, Ledford Hannah A, Lee Jeong Han, Chavez Karen S, Wang Wenying, López Javier E, Lieu Deborah K, Sirish Padmini, Zhang Xiao-Dong,



(8) Cationic graphene oxide nanoplatform mediates miR-101 delivery to promote apoptosis by regulating autophagy and stress.[TOP]

Pubmed ID :30319254
Publication Date : //
MicroRNA-101 (miR-101) is an intense cancer suppressor with special algorithm to target a wide range of pathways and genes which indicates the ability to regulate apoptosis, cellular stress, metastasis, autophagy, and tumor growth. Silencing of some genes such as with miR-101 can be interpreted as apoptotic accelerator and autophagy suppressor. It is hypothesized that hybrid microRNA (miRNA) delivery structures based on cationized graphene oxide (GO) could take superiority of targeting and photothermal therapy to suppress the cancer cells.

Authors : Assali Akram, Akhavan Omid, Mottaghitalab Fatemeh, Adeli Mohsen, Dinarvand Rassoul, Razzazan Shayan, Arefian Ehsan, Soleimani Masoud, Atyabi Fatemeh,



(9) Effects of HMGA2 gene downregulation by siRNA on lung carcinoma cell migration in A549 cell lines.[TOP]

Pubmed ID :30317663
Publication Date : //
Although there are multiple treatments for lung cancer, the death rate of this cancer remains high because of metastasis in earlier stages. So a novel treatment for overcoming metastasis is urgently needed. Overexpression of high-mobility group AT-hook 2 (HMGA2), a nonhistone chromosomal protein has been observed in metastatic cancers. So, we suggested that HMGA2 upregulation may play a critical role in treating lung cancer.

Authors : Naghizadeh Sanaz, Mansoori Behzad, Mohammadi Ali, Kafil Hossein Samadi, Mousavi Zohreh, Sakhinia Ebrahim, Baradaran Behzad,



(10) Mechanism of piR-DQ590027/MIR17HG regulating the permeability of glioma conditioned normal BBB.[TOP]

Pubmed ID :30305135
Publication Date : //
The blood-brain barrier (BBB) strongly restricts the entry of anti-glioma drugs into tumor tissues and thus decreases chemotherapy efficacy. Malignant gliomas are highly invasive tumours that use the perivascular space for invasion and co-opt existing vessels as satellite tumor form. Because regulation of the effect of noncoding RNA on BBB function is attracting growing attention, we investigated the effects of noncoding RNA on the permeability of glioma conditioned normal BBB and the mechanism involved using PIWI-associated RNA piR-DQ590027 as a starting point.

Authors : Leng Xue, Ma Jun, Liu Yunhui, Shen Shuyuan, Yu Hai, Zheng Jian, Liu Xiaobai, Liu Libo, Chen Jiajia, Zhao Lini, Ruan Xuelei, Xue Yixue,