Electroporation Therapy: A Transformative Approach to Enhancing Cellular Permeability.
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Most cited protocols related to «Electroporation Therapy»
All procedures were in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and approved by the Massachusetts Institute of Technology Committee on Animal Care. C57BL/6J E16-timed pregnant mice were used for electroporation. Surgery was done under ketamine-xylazine anesthesia and buprenorphine analgesia. For cortical experiments, DNA solution containing plasmids of interest were injected into lateral ventricle of each embryo using a pulled capillary tube. Five square pulses (50ms width, 1Hz, 35V) were applied using tweezer electrode for electroporation (Harvard Apparatus, ECM 830). Direct opsin-expressing experimental mice were electroporated with pCAG-opsin-GFP plasmid. Post-synaptic experimental mice were electroporated with pCAG-FLEX-rc[Chronos-GFP] and/or pCAG-FLEX-Chrimson-mOrange2, and pCAG-Cre plasmids. pCAG-Chrimson-tdTomato was additionally used in half of the single post-synaptic experiments. For the retinal ganglion cell-superior colliculus experiment, intravitreal virus injection was performed on P0 C57BL/6 mice with Nanoject II (Drummond) under cold anesthesia. 100 nL of rAAV2/8-Synapsin-Chronos-GFP (titer 1.4×1013 particles/mL) was injected into the eye. AAV particles were produced by the University of North Carolina Chapel Hill Vector Core.
Klapoetke N.C., Murata Y., Kim S.S., Pulver S.R., Birdsey-Benson A., Cho Y.K., Morimoto T.K., Chuong A.S., Carpenter E.J., Tian Z., Wang J., Xie Y., Yan Z., Zhang Y., Chow B.Y., Surek B., Melkonian M., Jayaraman V., Constantine-Paton M., Wong G.K, & Boyden E.S. (2014). Independent Optical Excitation of Distinct Neural Populations. Nature methods, 11(3), 338-346.
All procedures were in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and approved by the Massachusetts Institute of Technology Committee on Animal Care. C57BL/6J E16-timed pregnant mice were used for electroporation. Surgery was done under ketamine-xylazine anesthesia and buprenorphine analgesia. For cortical experiments, DNA solution containing plasmids of interest were injected into lateral ventricle of each embryo using a pulled capillary tube. Five square pulses (50ms width, 1Hz, 35V) were applied using tweezer electrode for electroporation (Harvard Apparatus, ECM 830). Direct opsin-expressing experimental mice were electroporated with pCAG-opsin-GFP plasmid. Post-synaptic experimental mice were electroporated with pCAG-FLEX-rc[Chronos-GFP] and/or pCAG-FLEX-Chrimson-mOrange2, and pCAG-Cre plasmids. pCAG-Chrimson-tdTomato was additionally used in half of the single post-synaptic experiments. For the retinal ganglion cell-superior colliculus experiment, intravitreal virus injection was performed on P0 C57BL/6 mice with Nanoject II (Drummond) under cold anesthesia. 100 nL of rAAV2/8-Synapsin-Chronos-GFP (titer 1.4×1013 particles/mL) was injected into the eye. AAV particles were produced by the University of North Carolina Chapel Hill Vector Core.
Klapoetke N.C., Murata Y., Kim S.S., Pulver S.R., Birdsey-Benson A., Cho Y.K., Morimoto T.K., Chuong A.S., Carpenter E.J., Tian Z., Wang J., Xie Y., Yan Z., Zhang Y., Chow B.Y., Surek B., Melkonian M., Jayaraman V., Constantine-Paton M., Wong G.K, & Boyden E.S. (2014). Independent Optical Excitation of Distinct Neural Populations. Nature methods, 11(3), 338-346.
Experiments were performed in accordance to the regulations of the IACUC at UCSD. Mice (except for Fig. 3a,b) were heterozygous for SOM-IRES-CRE (Jackson lab stock #013044) or PV-CRE (#008069) and the reporter allele Rosa-LSL-tdTOMATO (Allen Institute line Ai9, Jackson Labs #007905). For Fig. 3a,b mice were positive for Scnn1a-tg3-CRE (Jackson labs #009613) and crossed with to the Gin (#003718) or B13 line. For in vivo experiments mice were implanted with a custom head plate and habituated to head-fixation while running on a free spinning circular treadmill. For targeted recording in vivo tdTomato-expressing neurons were visualized by two photon microscopy and contacted by a glass electrode containing Alexfluor 488. Extracellular unit recording was performed via 16 channel silicon probes (Neuronexus). Single units were isolated using custom spike sorting software (Kleinfeld lab). We conditionally expressed ChR2 by in utero electroporation (for layer 2/3) or via a CRE-depednent AAV in Scnn1a-tg3-CRE (for layer 4). Arch or eNpHR were expressed via CRE-dependent AAVs in SOM- and PV-IRES-CRE mice. Visual stimuli were generated by custom software (Psych Toolbox) and presented on a gamma-corrected LCD monitor 15 cm from the mouse. Photostimulation in vivo was performed via fiber-coupled LEDs (Doric lenses). Photostimulation in vitro was via a combination of fiber-coupled LEDs, or LEDs mounted and coupled to an epifluorescence microscope (Olympus BX51). eNpHR was activated by a shuttered arc-lamp. Slice preparation and intracellular recording followed previous protocols. Data acquisition, visual stimulation, and statistical analysis was performed in the Igor Pro and Matlab environments.
Adesnik H., Bruns W., Taniguchi H., Huang Z.J, & Scanziani M. (2012). A Neural Circuit for Spatial Summation in Visual Cortex. Nature, 490(7419), 226-231.
Fluorescein-conjugated albumin protein is delivered in high concentration to the interior of 3T3 cells with high efficiency using protein electroporation, according to the method of Koken et al., 1994.
Protein Electroporation
3T3 cells electroporated at 300V with a varying number of 5 ms pulses showed progressively increased protein uptake (see FIGS. 4A-4D). ˜200,000 3T3 cells were electroporated in a 4 mm cuvette in the presence of FITC-conjugated albumin (150 ug/200 ul). Visual inspection and photography using a fluorescent microscope revealed progressively increased FITC-albumin uptake and fluorescence over a wide range of pulse number (10-200 pulses). FIGS. 4A-4D depict the cells (at low power magnification, 10×) 48 hours after exposure to 20 pulse (FIG. 4A), 50 pulse (FIG. 4B), 100 pulse (FIG. 4C) and 200 pulse (FIG. 4D) electroporation, demonstrating protein uptake was a function of electroporation.
US11859168B2. Electroporation, developmentally-activated cells, pluripotent-like cells, cell reprogramming and regenerative medicine (2024-01-02). None [US]. Inventors: Christopher B. Reid [US], Melissa Braga [US], Lilian N. Santamaria [US], Ashley N. Wickrema [US], Marinne D. Wickrema [US], Anthony Hao Dinh [US], Yaman Eksioglu [US], Mat Hoang Ho [US].
pKM225 was introduced into stationary phase C. burnetii NMII via electroporation. C. burnetii was made electrocompetent through washing with ice-cold, sterile filtered 10 % (v/v) glycerol prior to electroporation of 50 µl aliquots at 1.8 kV, 500 Ω and 25 µF using 0.1 cm cuvettes and an ECM630 electroporator (BTX Harvard Apparatus). Immediately after electroporation, 950 µl RPMI-glutamax was added to cuvettes. Five hundred microlitres of sample was used to inoculate 6 ml ACCM-2 medium in duplicate. Cultures were incubated overnight, before the addition of chloramphenicol at 3 µg ml−1, followed by a further 4 days of incubation. To obtain colonies, samples were plated onto ACCM-2 agarose plates containing chloramphenicol at 3 µg ml−1. After 14 days of incubation, colonies were washed from the plate and pooled. One hundred microlitres was used to inoculate triplicate 25 ml ACCM-2 cultures for TraDIS library preparation.
Metters G., Hemsley C., Norville I, & Titball R. (2023). Identification of essential genes in Coxiella burnetii. Microbial Genomics, 9(2), mgen000944.
Pregnant ICR mice were purchased from SLC Japan. Animals were handled in accordance with guidelines established by Keio University, Kyoto University, Tohoku Medical and Pharmaceutical University, FBRI and RIKEN‐BDR. All electroporations in this report were performed on E14 embryos. In utero electroporation experiments were performed as described previously with minor modifications (Kawauchi et al, 2003 (link)). Pregnant mice were deeply anesthetized and an abdominal or right dorsal incision was made to access the uterus. Approximately 1 μl of plasmid DNA (shRNA experiments: 3 μg/μl, low concentration of Ncad‐sh1023: 1 μg/μl, rescue experiments: 1–10 μg/μl, pCAG‐EGFP: 0.5 μg/μl) in endotoxin‐free TE buffer (Qiagen) containing Fast Green was injected into the lateral ventricle of embryonic brains with a glass micropipette (GD‐1, Narishige). Holding the embryo in utero with forceps‐type electrodes (NEPA GENE or BEX), 50 ms electric pulses of 35 V were delivered five times at intervals of 450 ms with a square electroporator (NEPA21, NEPA GENE or CUY21, BEX). After electroporation, the uterus was placed back into the abdominal cavity, allowing embryos to continue developing. At indicated stages, embryos were harvested and coronal sections of electroporated brains were prepared by using a cryostat (Leica).
Shikanai M., Ito S., Nishimura Y.V., Akagawa R., Fukuda M., Yuzaki M., Nabeshima Y, & Kawauchi T. (2023). Rab21 regulates caveolin‐1‐mediated endocytic trafficking to promote immature neurite pruning. EMBO Reports, 24(3), e54701.
For LUC complementation imaging assays53 (link), AtSYN4, AtSCC3, AtBMI1A/B/C, and AtRING1A were fused to the N- or C-terminal fragment of LUC (N and C), respectively. The primers used are listed in Supplementary Data 8. The fused plasmids were introduced into Agrobacterium tumefaciens strain GV3101 by electroporation and then incubated in LB (with 50 mg/L kanamycin and 25 mg/L gentamycin) plate medium at 30 °C for 48–72 h to OD600 = 0.8. Then mix the corresponding Agrobacterium tumefaciens strains equally and then co-infiltrated into tobacco (N. benthamiana) leaves using an injection syringe. 48 h later, the infiltrated leaves were injected with 100 mM luciferin (Sango, dissolved in water), and the luciferase signals were detected by the PMCapture software (Version 1.00) of a Chemiluminescence Imaging System (Tanon 5500, Shanghai, China). Values came from three biological replicates.
Zhang Y., Ma M., Liu M., Sun A., Zheng X., Liu K., Yin C., Li C., Jiang C., Tu X, & Fang Y. (2023). Histone H2A monoubiquitination marks are targeted to specific sites by cohesin subunits in Arabidopsis. Nature Communications, 14, 1209.
The biotinylation efficiency of EVs and the optimization of electroporation were analyzed by using an Apogee A‐50 Micro Flow Cytometer (Apogee Flow Systems) equipped with 375, 405, 488, and 638 nm lasers. The reference beads (ApogeeMix, Apogee Flow Systems) composed of a mixture of nonfluorescent silica beads (Si, refractive index ≈1.42) with diameters of 180, 240, 300, 590, 880, 1300, and 110 nm, 500 nm green fluorescent latex spheres (polystyrene, refractive index ≈1.59) were used to set the thresholds for light scattering and help to gate sEVs and lEVs. The tubing was washed with double‐distilled water after each sampling. Analysis was performed at a flow rate of 1.5 µL min−1 using a 150 µL sample volume for at least total 3 × 105 particles.
Xia H., Yu Z., Zhang L., Liu S., Zhao Y., Huang J., Fu D., Xie Q., Liu H., Zhang Z., Zhao Y., Wu M., Zhang W., Pang D, & Chen G. (2023). Real‐Time Dissection of the Transportation and miRNA‐Release Dynamics of Small Extracellular Vesicles. Advanced Science, 10(7), 2205566.
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Fast Green is a laboratory staining dye used in various scientific applications. It is a synthetic, water-soluble dye that provides a green coloration. The core function of Fast Green is to stain and visualize specific components or structures within biological samples during microscopy and other analytical procedures.
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Opti-MEM is a cell culture medium designed to support the growth and maintenance of a variety of cell lines. It is a serum-reduced formulation that helps to reduce the amount of serum required for cell culture, while still providing the necessary nutrients and growth factors for cell proliferation.
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The Neon Transfection System is a laboratory equipment designed for the efficient delivery of nucleic acids, such as DNA or RNA, into a variety of cell types. It utilizes electroporation technology to facilitate the transfer of these molecules across the cell membrane, enabling researchers to study gene expression, conduct functional assays, or modify cellular behavior.
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The Gene Pulser Xcell is a laboratory instrument designed for electroporation, a technique used to introduce genetic material into cells. It generates an electrical pulse that temporarily creates pores in the cell membrane, allowing foreign DNA or RNA to enter the cells. The device can be used for a variety of cell types, including bacterial, plant, and mammalian cells.
The ECM 830 is a single-channel, computer-controlled electrophysiology stimulator designed for tissue culture and in vitro studies. It provides programmable electrical stimulation with adjustable voltage, current, and pulse duration parameters. The device is capable of delivering both monophasic and biphasic stimuli to biological samples.
The Gene Pulser Xcell Electroporation System is a laboratory instrument designed for the delivery of nucleic acids, such as DNA or RNA, into cells through the process of electroporation. The system provides precise control over the electroporation parameters, allowing users to optimize the efficiency of gene transfer.
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The Gene Pulser is a laboratory equipment designed for electroporation, a technique used to introduce genetic material into cells. It generates an electrical pulse that temporarily increases the permeability of cell membranes, allowing for the uptake of DNA, RNA, or other molecules. The core function of the Gene Pulser is to facilitate this electroporation process in a controlled and reliable manner.
The NEPA21 Super Electroporator is a laboratory instrument designed for the efficient transfer of genetic material, such as DNA or RNA, into cells. It utilizes electrical pulses to temporarily increase the permeability of cell membranes, allowing the introduction of the desired genetic material. The device features adjustable voltage and pulse duration settings to optimize the electroporation process for a variety of cell types.
The NEPA21 is a laboratory equipment designed for electroporating cells. It utilizes programmable pulse settings to facilitate efficient DNA, RNA, or protein delivery into cells.
The CUY650P5 is a laboratory equipment product designed for scientific research applications. It serves as a platform for conducting various experiments and analyses. The core function of the CUY650P5 is to provide a controlled environment for sample preparation, observation, and data collection.
Electroporation Therapy is a transformative approach that enhances cellular permeability, allowing for the delivery of various molecules, including drugs, genes, and other therapeutics, into cells. This non-invasive technique utilizes controlled electrical pulses to create temporary pores in the cell membrane, facilitating the uptake of these important compounds.
Electroporation Therapy encompasses several variations, including in vivo, ex vivo, and in vitro applications. Each type has its own unique set of parameters and considerations, such as pulse duration, voltage, and electrode configuration, which can be optimized for specific research or clinical needs.
Electroporation Therapy has a wide range of applications, including cancer treatment, gene delivery, vaccine development, and tissue engineering. It has been used to enhance the delivery of chemotherapeutic agents, introduce genetic material for gene therapy, and facilitate the incorporation of biomaterials in tissue regeneration.
One of the key challenges with Electroporation Therapy is ensuring optimal protocol selection and reproducibility across different research settings. Factors such as cell type, tissue characteristics, and electrical parameters can significantly impact the efficacy and safety of the technique. Additionally, proper optimization and standardization of the process are crucial to achieving consistent and reliable results.
PubCompare.ai's AI-driven research protocol comparison tool can greatly assist with Electroporation Therapy research. The platform allows you to efficiently screen protocol literature, leveraging AI to pinpoint critical insights. PubCompare.ai can help researchers identify the most effective protocols related to Electroporation Therapy for their specific research goals, highlighting key differences in protocol effectiveness and enabling them to choose the best option for reproducibility and accuracy. This can streamline the research process and unlock new possibilities in this dynamic field.
More about "Electroporation Therapy"
Electroporation therapy (EP) is a revolutionary approach that enhances cellular permeability, enabling the effective delivery of a variety of molecules, including DNA, RNA, proteins, and small molecules, into cells.
This technique utilizes brief, high-voltage electrical pulses to create temporary pores in the cell membrane, facilitating the entry of therapeutic agents.
Discover how PubCompare.ai's AI-driven research protocol comparison can optimize your electroporation therapy journey.
This innovative platform allows you to locate the best electroporation protocols from the literature, preprints, and patents, ensuring you have access to the most up-to-date and effective techniques.
By enhancing reproducibility and accuracy with PubCompare.ai's intelligent analysis tool, you can streamline your research and unlock new possibilities in this dynamic field.
Explore the various electroporation systems available, such as the Neon Transfection System, Gene Pulser Xcell, ECM 830, and NEPA21 Super Electroporator.
These cutting-edge technologies offer a range of features and capabilities to suit your specific needs, from high-throughput applications to precise control over pulse parameters.
In addition to electroporation, consider the use of complementary techniques like Fast Green and Opti-MEM to optimize your cellular delivery and enhance your research outcomes.
By leveraging the synergistic effects of these methods, you can maximize the efficiency and effectiveness of your electroporation therapies.
Take your electroporation therapies to new heights by exploring PubCompare.ai today.
Discover how this AI-powered platform can streamline your research, improve reproducibility, and unlock new possibilities in this rapidly evolving field.