CRO服务

Disease-related gene stable strain construction

Basic Information

　　Stable Transgenic Strains refers to a cell line obtained stable overexpression or inhibition of a specific gene through gene editing. By means of loading disease-related gene sequences or shRNA sequences targeting specific genes onto a recombinant vector, and integrating the target gene sequence into the chromosome of the host cell through a lentivirus screening system or a transposon screening system, it can achieve sustained and stable expression and regulation of the target gene. Stable Transgenic Strains include overexpressing stable cell lines, knockdown stable cell lines, and disease-specific gene stable cell lines.

 Services We Provide

　　We can generate stable cell lines through the lentiviral drug screening system (Lentivirus Infection-Drug Screening Method) or a transposon screening system, which includes the construction of overexpression stable cell lines, knockdown stable cell lines, or disease-associated gene stable cell lines.

Procedure of Services

1.Lentivirus Screening System
（1）Target Gene Screening
　　The target genes or indication-specific target genes are selected by bioinformatics analysis.
（2）Cell Line Selection
　　The cell lines selected should be consistent with the indications, for example, when studying hepatocellular carcinoma, we should select corresponding hepatocellular carcinoma cell lines for stable cell line construction, which can be also used for downstream in vivo anti-tumor activity evaluation. QPCR and Western blot are used to detect the gene expression in cell lines, and the appropriate cell line for stable target gene construction are selected based on the gene expression.
　　In addition, if studying the interaction between drugs and targets, the tool cell lines such as 293T and CHO-K1 are recommended. These tool cell lines are easy to be infected by lentivirus, which facilitates the generation of stable cell lines.
（3）Vector Construction
　　Load the coding sequence (CDS) of the target gene or the shRNA sequence that inhibits target gene into the recombinant vector.
（4）Lentiviral packaging and plasmid extraction.
（5）Lentiviral Packaging
　　Co-transfecting lentiviral plasmids into 293T cells to generate lentiviral particles via a lentiviral packaging system.
（6）Lentiviral Infection
　　Set different gradient MOIs (multiplicity of infection) to screen for the optimal MOI.
（7）Antibiotic Screening
　　Common resistance selection markers for eukaryotic expression vectors include neomycin, hygromycin, puromycin, and G418 Geneticin. Different antibiotics are selected based on the specific resistance sequences to conduct drug screening on cells.
（8）The Construction of Polyclonal/Monoclonal Stable Cell Lines
（9）Validation of Stable Transgenic Cell line Expression
　　The expression of target gene was detected by qPCR or Western blotting assays.
（10） 交付实物Delivery of Experimental Results
　　T25 cell flask or cryovial containing 1*106 of cells or 2*106 of cells.


2.Transposon Screening System
（1）Target gene screening
　　Target genes or target genes for indications are screened by bioinformatics.
（2）Construction of vectors and plasmid extraction.
（3）Co-transformation of transposase plasmid and transposon sequence plasmid into cells
（4）Antibiotic screening
　　Common resistance selection markers for eukaryotic expression vectors include neomycin, hygromycin, puromycin, and G418 Geneticin. Different antibiotics are selected based on the specific resistance sequences to conduct drug screening on cells.
（5）Monoclonal amplification
　　Expanding and culturing cells through monoclonal amplification
（6）Detecting whether the target gene sequence has been transposed into the genome.

Strengths and Features

（1）One stop integrated services
　　with over 800 strains of cell lines in our cell bank, we have complete resources of seed stocks and provide end-to-end services for your research.
（2）Professional technical support
　　Jennio has successfully constructed over 500 stable transgenic cell lines through the "Lentivirus Infection-Drug Screening Method" and these cell lines maintain stable expression after passage.
（3）Multiple stable transgenic strain screening systems are available for selection.
（4） On-time delivery
　　It takes approximately 3-4 months from plasmid construction to delivery of multi-clone stable transgenic strains, and about 4-6 months from plasmid construction to delivery of monoclonal stable transgenic strains.
（5） Quality assurance
　　We ensure that our cell strains are free from mycoplasma and chlamydia infections through strict quality control measures.
（6）Professional technical guidance
　　Our technical team has an average experience of more than 6 years, and we can provide professional guidance on cell culture techniques.
（7） After-sales service
　　We provide a two-week no-obligation after-sales service to address any concerns you may have.

Representative Case

Our Laboratory