With the improvement of Creative BioMart's technology platform, we already have the ability to provide stable cell line production services with the required genetic modification at a satisfactory speed and cost.
In addition to some regular vector-mediated cell lines that can stably express foreign proteins, we can also establish the
We can design and generate custom vectors to construct stable cell lines with specific genetic modifications. The final product will be delivered in the form of pooled cells or single clone.
Establish overexpression stable cell ine
Creative BioMart's overexpression cell lines are established based on lentiviral transduction. After the establishment of cell lines that stably overexpress your GOI, we will verify the expression level by RT qPCR. Finally, the expressed product will be detected by a series of standard QC analyses. You can contact us to understand the specific content of QC analyses.
Establish shRNA knockdown stable cell lines
The establishment of shRNA knockdown stable cell lines is still based on lentivirus packaging and expression technology. We will construct several lentivirus vectors that carry shRNA. The top 3 lentiviruses with the highest knockdown score will be selected to transfect target cells. The cell line that performs best in the RT-qPCR test will be selected. After QC testing, the cell pool or single clone will be delivered.
Tet inducible gene expression
Creative BioMart can also establish Tet inducible gene expression stable cell lines with minimal background expression and high induction of your GOI(s). Additionally, we can generate Tet regulatory protein expression stable cell lines (e.g. rtTA, tTS/rtTA, Tet3G, etc.) which can then be transfected/transduced with plasmids/viruses carrying TRE driven GOI(s) for flexible experimental design and reliable Tet inducible gene expression. For Tet-On stable cell lines, induction of the GOI is validated by RT-qPCR.
Gene knockout
We can generate stable cell lines with permanent knockout of your GOI(s). Stable knockout of GOI(s) is achieved using a CRISPR-based approach, wherein a pair of gRNAs targeting the GOI(s) is introduced into the target cells along with Cas9. This leads to the generation of two cuts on the target gene. Attempts by cells to repair the broken ends of the two cut sites would typically lead to a large deletion spanning the two sites. The two cut sites are designed to flank a functionally important region of the target gene such that a deletion spanning them would most likely lead to the loss of gene function.
Depending on the cell type, either lentiviral or regular plasmid CRISPR vectors may be used for achieving target gene knockout. The final cell line is evaluated by Sanger sequencing of the targeted locus to confirm knockout.
Gene knockin
We can generate stable cells line with permanent knockin of your GOI(s) at desired genomic target sites. Stable knockin of GOI(s) is achieved by a CRISPR-based approach wherein a target site-specific gRNA is introduced into the cells along with Cas9 and a donor vector which serves as the template for HDR-mediated gene knockin.
Point mutation
Creative BioMart can generate stable cell lines harboring desired point mutations at genomic target sites of interest. Stable cell lines with point mutations are generated using a CRISPR-based approach wherein a target site-specific gRNA is introduced into the cells along with Cas9 and a single-stranded donor oligonucleotide (ssODN) which serves as the template for HDR-mediated insertion of the point mutation.
The final cell line is evaluated by Sanger sequencing of the targeted locus to confirm the presence of the point mutation.
Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP. If you have any special needs or are interested in learning more about Creative BioMart's protein expression services, please feel free to inquire about more details.
