Lentiviral delivery of cDNAs (ORFs), shRNA and sgRNAs is widely used efficient approach to incorporate the desired genetic modifications in the cells of interest.
FINGEEC provides stable cell line generation services for research purposes. We use recombinant lentiviruses or retroviruses for transgene expression in human and other mammalian cell lines. Our stable cell line services are best suited for researchers who do not have access to a biosafety level 2 facility, have limited time for new technique testing or for researchers who are new to transgene technology.
Our cell line generation consultation services are also free of cost.
The inception of translational CRISPR (Clustered regularly interspaced short palindromic repeats) technology was in 2013, leading to Nobel Prizes for Jennifer Doudna and Emmanuelle Charpentier in 2020. Since then, CRISPR technology has been widely used for genetic manipulations in all kinds of systems with over 32,000 scientific publications utilizing the technology in one way or another.
At FinGEEC, we use CRISPR/Cas9 technology to create genetic editing in different mammalian cell lines for research purposes.
While the successful editing with the CRISPR/Cas9 depends on several factors, we have been able to generate cell lines (>25 target cell lines) with desired manipulations (almost 30 different genes) (mostly knockouts; check our list of genes and cell lines edited by FinGEEC) in humans and other mammalian cell lines.
Genome editing by CRISPR/Cas9 system is a popular laboratory technique used in introducing changes in the DNA sequences of genome. The editing event is based on a double strand break caused by a Cas9 nuclease, the specificity of which is defined by a guiding sequence of RNA that is complementary to desired site in the organism’s genome. Success of the CRISPR/Cas9 genome editing is dependent on many steps, but the use of well-designed and in vitro validated guide RNA constructs is instrumental for a successful genome editing project.
FINGEEC provides validation of the customer designed or commercial gRNAs in HEK293FT cells prior to generating CRISPR/Cas9 knockout cell lines.
We check the transfection efficacy of the constructs with fluorescence microscopy and assess the functionality of the gRNA with a T7 nuclease mutation detection assay. The gRNA functionality report makes it easy for researchers to choose the best gRNAs that they want to use in cell line generation.
The research infrastructure is supported by the University of Helsinki (HiLIFE and Research Programs Unit) and Biocenter Finland.
The prices are for the University of Helsinki (UH) and Universities/Institutions affiliated with Biocenter Finland (BF) Network; for other academic and non-academic customers, please contact fingeec-support@helsinki.fi or topi.tervonen@helsinki.fi
CELL-BASED SERVICES |
FEE (€) University of Helsinki |
fee (€) Biocenter finland |
---|---|---|
Stable cell line (1 ctrl line + 1 target line) | 479 | 594 |
Stable cell line (Extra order/line) | 243 | 301 |
gRNA validation for CRISPR guide RNA (1 ctrl guide RNA + 1 on-target guide RNA) | 215 | 265 |
gRNA validation for CRISPR guide RNA (Extra order/guide RNA) | 70 | 87 |
CRISPR cell line (Transient transfection) (1 ctrl line + 1 on-target KO line) | 541 | 671 |
CRISPR cell line (Transient transfection) (Extra order/on-target KO line) | 221 | 274 |
CRISPR cell line (Lentiviral delivery) (1 ctrl line + 1 on-target KO line) | 585 | 726 |
CRISPR cell line (Lentiviral delivery) (Extra order/on-target KO line) | 287 | 355 |
The success of the cell line generation project depends on cell line, target gene, the downstream effects of the target gene manipulation (especially those related to cell viability and proliferation), delivery methods and other factors. Below are some of the points to consider before starting a cell line project with FinGEEC:
Finnish Genome Editing Center (FinGEEC) should be mentioned in the acknowledgment in research publications containing data from cell lines generated by FinGEEC. Below is a template sentence that can be used:
For stable and CRISPR cell lines:
“The xx cell line/pool was generated by Finnish Genome Editing Center (FinGEEC), Research Programs Unit, Faculty of Medicine, University of Helsinki, supported by HiLIFE, University of Helsinki, and Biocenter Finland."