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Electroporation
Achieving efficient delivery of gene editing systems into cells is a crucial step for successful gene editing, as delivery efficiency is positively correlated with editing efficiency. Electroporation is a highly efficient delivery method that uses high-voltage electric pulses to create pores in the cell membrane, allowing the introduction of foreign DNA, RNA, or other molecules into the cells. This transfection method can be applied to most cell types and offers advantages such as high transfection efficiency and broad applicability.
Service Details
| Advantages of Electroporation | 1. High delivery efficiency 2. Broad cell applicability |
|---|---|
| Turnaround/Price |
 Consult online for details |
EDITGENE has developed a well-established electroporation platform, equipped with advanced instruments and optimized electroporation protocols. Leveraging years of experience in gene editing, we have created an efficient and comprehensive electroporation service system. Through this method, we can effectively deliver the editing system into target cells, followed by a selection process to obtain the desired cell lines.
EDI-Service Advantages
High Transfection Efficiency
Efficiently delivers plasmids or RNP complexes instantly.
Rapid Transfection of Large Quantities of Cells
Able to transfect large quantities of cells in a short period.
Wide Cell Applicability
Suitable for a wide range of cell types.
Mature Electroporation Platform
Equipped with professional instruments, protocols, and technical personnel.
Workflow
Case Study
Using the electroporation method, the HES-KI system was delivered into K562 cells, successfully integrating and expressing EGFP at the Safety Harbor locus.
1）Method
2）Result
Electroporation of K562 Cells with HES-KI System Without Selection to Generate Cell Pool
Literature Review
Traditional generation of gene-edited animals relies on microinjection of CRISPR/Cas9 proteins and donor DNA into zygotes, a method known for its low efficiency, time-consuming nature, and high technical skill requirements. Remy et al. reported an optimized electroporation method for delivering sgRNA and Cas9 proteins, with or without ssODN (single-stranded DNA fragments), into intact rat zygotes to improve gene-editing efficiency. Using ssODN as the donor template, this approach achieved precise knock-in mutations in 25-100% of the analyzed animals.
CRISPR/Cas9-Mediated HDR Strategy Based on ssODN
Remy, S., Chenouard, V., Tesson, L. et al. Generation of gene-edited rats by delivery of CRISPR/Cas9 protein and donor DNA into intact zygotes using electroporation. Sci Rep 7, 16554 (2017). https://doi.org/10.1038/s41598-017-16328-y.
Advantage and Characteristic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
FAQ
How to choose the appropriate gene delivery method?
Selecting a suitable gene delivery system requires a comprehensive assessment based on specific experimental conditions, research objectives, and cell types. Quantitatively comparing various systems in terms of delivery efficiency, cytotoxicity, and stability is an important step in determining the choice.
Viral delivery systems are suitable for experiments that require high delivery efficiency and sustained gene expression, especially when cells can tolerate higher levels of cytotoxicity and immune responses. If lower cytotoxicity and immune response, along with ease of use and cost-effectiveness, are priorities, then a liposome-based gene delivery system should be chosen. For high delivery efficiency that involves delivering large DNA fragments, and if the user can accept a higher operational complexity, a gene gun delivery system is an optional method. If high delivery efficiency is needed while maintaining relative simplicity and no special equipment is required, then the electroporation delivery system may be a suitable choice.
Viral delivery systems are suitable for experiments that require high delivery efficiency and sustained gene expression, especially when cells can tolerate higher levels of cytotoxicity and immune responses. If lower cytotoxicity and immune response, along with ease of use and cost-effectiveness, are priorities, then a liposome-based gene delivery system should be chosen. For high delivery efficiency that involves delivering large DNA fragments, and if the user can accept a higher operational complexity, a gene gun delivery system is an optional method. If high delivery efficiency is needed while maintaining relative simplicity and no special equipment is required, then the electroporation delivery system may be a suitable choice.
What issues should be considered when culturing cells for gene delivery?
Maintaining the activity of cell cultures is crucial. Cells should not be allowed to reach confluence for more than 24 hours. Frozen new cells can restore transfection activity. The optimal cell plating density varies for different cell types or applications; however, for adherent cells, a confluence of 70% to 90% or a density of 5×10^5 to 2×10^6 suspended cells/ml typically yields good transfection results. It is important to ensure that cells are not fully confluent or in a fixed phase during transfection.
