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Stable expression cell lines refer to the consistent and stable expression of a specific gene or interference with the expression of a specific gene in a cell. The plasmid DNA of the target gene of a stable cell line is integrated into the cell chromosome so that the cell consistently expresses the gene or consistently interferes with the target gene over a long period of time. BOC Sciences has expertise in stable cell line construction and we can perform RNAi stable cell line screening services as required by our clients.
What is RNAi Stable Cell Line Screening?
Stable transfection is the integration of the transfected plasmid DNA into the chromosome of the host cell so that the host cell can express the target gene and protein for a long time. It is necessary to screen target cells based on transient transfection or use high-transfection-efficiency viruses, and select corresponding drugs according to the resistance markers contained in different gene carriers for cell passage, to obtain cell lines that can stably express the target gene.
Stable cell line screening methods:
- After transfection of the plasmid, the monoclonal method of screening stable cell lines is less efficient for the transfection of most cells. For gene overexpression, if the transfection efficiency reaches 40%, gene overexpression is still possible. However, for gene interference experiments, the requirement for transfection efficiency is much higher than that for gene overexpression, which usually cannot be met by plasmid transfection. In addition, the plasmids are free in the cytoplasm and degrade quickly, making them unsuitable for longer-term assays. The probability of plasmid transfection and integration is extremely low, and the construction of stable cell lines is time-consuming and labor-intensive, and the yield is very low, so it is often necessary to pick a single clone.
- Viral infection screening of stable cell lines: The viral infection method is more convenient and efficient than the plasmid transfection screening of monoclonal methods, and is currently the mainstream method for screening stable cell lines. The use of lentivirus for the preparation of stable cell lines is an ideal vector for the preparation of stable cell lines due to its efficient integration, efficient transcription, efficient expression, wide host range, high infection efficiency and integration with cell chromosomes without gene rearrangement.
Our RNAi Stable Cell Line Screening Service
- Antibiotic screening concentration determination (lethality curve) - take the antibiotic concentration at which all cells die in 10-14 days as the screening concentration.
- Cell inoculation - cells are inoculated the day before the transfection experiment, and the plate density of each cell is determined by the growth rate and cell shape of each cell. On the day of transfection, the cell density should be 60% to 80% coverage.
- Cell transfection (liposome) or infection (lentivirus).
- Screening for stable interfering cell lines using plasmid resistance.
- validation of target gene interference effect.
Advantages of RNAi Stable Cell Line Screening Services from BOC Sciences
- Mature technology platform - rich experience in gene editing, with a mature and stable platform.
- First-class expert technical team - the team is professional and experienced.
- Reasonable prices - affordable and cost effective.
- Quality assurance - rigorous identification of exogenous genes.
- Professional after-sales service - a professional after-sales team is in place to answer and receive after-sales calls from customers, providing practical solutions for customers efficiently and quickly.
* Only for research. Not suitable for any diagnostic or therapeutic use.
