Exo fect sirna mirna transfection kit

Manufactured by System Biosciences

Sourced in United States

The Exo-Fect siRNA/miRNA Transfection Kit is a product designed for the efficient delivery of small interfering RNA (siRNA) or microRNA (miRNA) into cells. It is a transfection reagent that facilitates the introduction of these genetic materials into the target cells.

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Lab products found in correlation

Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (3 mentions) Hiperfect transfection reagent, by Qiagen (1 mentions) Laser confocal microscope, by Leica (1 mentions) Dapi blue, by Beyotime (1 mentions) Balb c nude mice, by Charles River Laboratories (1 mentions) Te 3000, by Nikon (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions) Nextseq 500, by Illumina (1 mentions) Mirneasy mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

Exo-Fect siRNA/miRNA Transfection Reagent kit Exo-Fect

7 protocols using exo fect sirna mirna transfection kit

Modulating EV miRNA and VSMC PiT-1 in Experiments

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Isolated EV^UR/EV^CTRL were transfected with miR inhibitors, miR mimics or respective controls using the Exo‐Fect™ siRNA/miRNA Transfection Kit (System Biosciences, Palo Alto, CA, USA) according to the manufacturer's instructions. All used miR inhibitors and miR mimics are given in Table S2. 24 hours after transfection, the EV were used in the respective experiments.
VSMC were transfected with siRNA targeted against PiT‐1 (Silencer™ Slc20a1 siRNA, #AM16708, Invitrogen™) or a respective negative control (Silencer™ Negative Control No. 1 siRNA, #AM4611, Invitrogen™) using the HiPerFect Transfection Reagent (Qiagen) according to the manufacturer's instructions.

Freise C., Querfeld U., Ludwig A., Hamm B., Schnorr J, & Taupitz M. (2021). Uraemic extracellular vesicles augment osteogenic transdifferentiation of vascular smooth muscle cells via enhanced AKT signalling and PiT‐1 expression. Journal of Cellular and Molecular Medicine, 25(12), 5602-5614.

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Modulating latent HIV with pEVs

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pEVs were transfected with miR-139-5p inhibitor (500 nM) using the Exo-fect™ siRNA/miRNA transfection kit (System Biosciences) according to manufacturer’s instructions. Transfected pEVs (HC, HIV naïve and ART) were applied to J-Lat 10.6 cells. Cells were washed after overnight incubation, fresh media added and harvested after 72hrs. Mock transfected pEVs were used as controls.

Okoye I., Xu L., Oyegbami O., Shahbaz S., Pink D., Gao P., Sun X, & Elahi S. (2021). Plasma Extracellular Vesicles Enhance HIV-1 Infection of Activated CD4+ T Cells and Promote the Activation of Latently Infected J-Lat10.6 Cells via miR-139-5p Transfer. Frontiers in Immunology, 12, 697604.

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Exosomal miR-148a-3p Uptake and Analysis

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Cy5-labelled hsa-mir-148a-3p mimic and mimic control were purchased from RiboBio (Guangzhou, China). Briefly, Exosomes extracted by the ExoQuick-TC were resuspended in 300μL PBS and transfected with miRNA mimics using Exo-Fect siRNA/miRNA Transfection Kit (System Biosciences, Palo Alto, CA) according to the manufacturer's protocol.
For exosome-uptaking assay, transfected exosomes were labelled with PKH67 (Sigma-Aldric, St. Louis, MO, USA) away from light at room temperature for 4min. After incubation with pre-treated exosomes for 24h, tumor cells were stained with phalloidin-rhodamine (Red, Sigma-Aldric) and DAPI (Blue, Beyotime). Images of tumor cells were captured using a laser confocal microscope (Leica, Germany) and qRT-PCR was performed to validate the miRNA expression level.

Zhang X., Chen F., Huang P., Wang X., Zhou K., Zhou C., Yu L., Peng Y., Fan J., Zhou J., Lu Z., Hu J, & Wang Z. (2022). Exosome-depleted MiR-148a-3p derived from Hepatic Stellate Cells Promotes Tumor Progression via ITGA5/PI3K/Akt Axis in Hepatocellular Carcinoma. International Journal of Biological Sciences, 18(6), 2249-2260.

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Exosomal miRNA Modulation in Hepatoma

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To investigate the roles of exosomes derived from HSCs, HCCLM3 cells transfected with luciferase lentivirus (1×10⁷) were inoculated subcutaneously into the right armpit of 4-week-old female Balb/c nude mice (Charles River laboratories, Beijing, China) housed in laminar-flow cabinets under specific pathogen-free conditions. After euthanasia of animals, tumors were procured and cut into small pieces of approximately 2 mm³. Subsequently, in situ hepatoma murine models were generated by implanting tumor pieces into the left lobe of liver. Exosomes (10 ug) extracted from the cell culture media of LX-2 were transfected with 1nmol Cy5- labelled hsa-mir-148a-3p agomir (Ribobio, Guangzhou, China) or negative control using Exo-Fect siRNA/miRNA Transfection Kit (System Biosciences, Palo Alto, CA) prior to injecting into mice through tail vein. The exosomes treatment started on day 7 and was performed every three days for four weeks. Then, all mice were euthanized and tumor tissues were cryo-sectioned and processed for fluorescent detection. All related experimental protocols were approved by the Institutional Animal Care and Use Committee at Zhongshan Hospital, Fudan University (Shanghai, China).

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Exosomal delivery of NANPs

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100 μg of isolated exosomes were loaded with 25 pmoles NANPs using the Exo-Fect siRNA/miRNA transfection kit (System Biosciences, Palo Alto, CA, USA) according to the manufacturer’s manual. The loaded exosomes were cleaned to remove any excess of free NANPs/negative control siRNA, transfection reagent, or complexes. The cleaned NANP loaded exosomes were immediately added onto MDA-MB-231-GFP cells.

Nordmeier S., Ke W., Afonin K.A, & Portnoy V. (2020). Exosome mediated delivery of functional nucleic acid nanoparticles (NANPs). Nanomedicine : nanotechnology, biology, and medicine, 30, 102285.

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Uptake of Cy3-Tagged Extracellular Vesicles by HMVECs

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Uptake of ssEVs by HMVECs was examined by treating the HMVECs with Cy3‐tagged ssEVs. The ssEVs were tagged with Cy3 by loading Cy3 siRNA using Exo‐Fect siRNA/miRNA Transfection Kit (Cat. #: EXFT200A‐1, System Biosciences) according to the manufacturer's protocol. Then HMVECs were treated with Cy3‐tagged ssEVs (10⁸ particles/mL) for 48 hours. Uptake of Cy3‐tagged ssEVs was evaluated using a fluorescent microscope (Nikon TE3000).

Cheng Z., Naga Srikanth Garikipati V., Truongcao M.M., Cimini M., Huang G., Wang C., Benedict C., Gonzalez C., Mallaredy V., Goukassian D.A., Verma S.K, & Kishore R. (2021). Serum‐Derived Small Extracellular Vesicles From Diabetic Mice Impair Angiogenic Property of Microvascular Endothelial Cells: Role of EZH2. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 10(10), e019755.

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Isolation and Analysis of iMSC-sEV miRNAs

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iMSC-sEV miRNAs were isolated using an miRNeasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s protocol. The concentration and purity of the RNA samples were detected using a NanoDropND-1000 spectrophotometer (Thermo Scientific, Waltham, MA, USA). Microarray analysis was performed on an Illumina NextSeq 500 (Illumina, San Diego, CA, USA) and analyzed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA).
We searched the PubMed database for microRNAs (miRNAs), involved in promoting either NSC proliferation or neuronal differentiation using the search terms: ([NSC* or neural stem* or neural progenitor* or NPC*]) AND ([miRNA* or microRNA*]]) AND ([neurogenesis* or proliferation*]). Searches were limited to English language articles.
miRNA inhibitors of miR-21-5p and miR-486-5p and a negative control were obtained from GenePharma (Shanghai, China). All the nucleotides in the inhibitors contained 2′-O-Me modifications at every base and a 5′-Cy3-containing amino linker. The sequences of the inhibitors are listed in Table 1. An Exo-Fect™ siRNA/miRNA Transfection Kit (System Biosciences, Mountain View, CA, USA) was used to deliver the miRNA inhibitors into the iMSC-sEVs in accordance with the manufacturer’s instructions.

Lang H.L., Zhao Y.Z., Xiao R.J., Sun J., Chen Y., Hu G.W, & Xu G.H. (2022). Small extracellular vesicles secreted by induced pluripotent stem cell-derived mesenchymal stem cells improve postoperative cognitive dysfunction in mice with diabetes. Neural Regeneration Research, 18(3), 609-617.

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