Alpha Lifetech Inc. is experienced in cell line construction. We are committed to providing high-quality stable cell line construction services to customers worldwide and customized services are available to support your research. Our services are proven and trusted.
Stable Cell lines Construction - Low Expression of Stable Cell lines
RNA interference technology is used to reduce the expression level of target proteins. Generally, we don't want to knock out a gene in a stable cell line, but we need to reduce the expression level of protein of interest in it, that's why gene silencing technology, namely RNA interference technology we need. With many years of experience in genetic engineering, scientists in Alpha Lifetech Inc. have developed many stable cell lines with genes deleted (gene knock-out), or partly gene silencing (gene knock down) and gene over-expression (gene knock in).
RNA interference (RNAi) often used to reduce the expression of proteins of interest in stable cell line. RNAi is the process by which expression of a target gene is effectively silenced or knocked down by the selective inactivation of its corresponding mRNA by double-stranded RNA (dsRNA). RNAi is activated by dsRNA species delivered to the cytoplasm of cells. The silencing mechanisms can either lead to the degradation of a target mRNA, as induced by small interfering RNAs (siRNAs) or short hairpin RNAs (shRNAs), or the suppression of translation of specific mRNAs, as induced by microRNA (miRNA).
Gene knockdown often uses the lentiviral expression system as a delivery method, in which U6 promoter-driven pLVshRNA-puro and pLVshRNA-EGFP (2A) plasmid, as carriers of target genes, are used to transfect with the cell line for screening cells with immortalized low expression of protein of interest. The schematic model of pLVshRNA-EGFP Plasmid shows as bellow:
The pLVshRNA-eGFP vector was designed to interfere with the expression of the target gene through shRNA synthesis in host cells. RNAi sequence has been cloned into multiple clone sites (MCS), and the type III RNA polymerase uses the U6 promoter to transcript shRNA which finally resulting in the interference effect of gene of interest. EGFP fluorescent protein encoded by pLVshRNA-eGFP vector, can be used as a signal to observe the efficiency of virus infection and to screen positive cells by flow cytometry.
Service Features of Low Expression of Stable Cell lines
-- High Success Rate: 100% Guarantee
-- Stable Gene Integration
-- High Interference Efficiency: RT-PCR Validation (> 80%)
-- High-quality Polyclonal Cell Lines or Monoclonal Cell Lines
-- Rigorously validated cell lines for accurate pharmacology
-- Single assay format for detection in agonist, antagonist, inverse agonist, and allosteric modulator modes
-- Chemiluminescent detection for fast and reliable results, suitable for automation
-- Broad applications ideal for small molecule and biologics screening or profiling
If you have any questions or would like to receive more information about this program, please feel free to contact us at any time. Our scientists would be glad to hear from you and discuss with your project.