Virus vectors

AAV titration

The titration measures the number of viral genomes/ml, using the qPCR method. This is based on a comparison to a standard viral sample of known titer. The customer should provide 10 to 100 µl of cell culture medium or 2 µl of a concentrated virus.

Price for academic users: Basic fee: 68 € +7 €/ sample

Lentivirus titration

We measure the number of viral genomes per ml of the provided sample (10 to 100 µl cell culture supernatant or 2 µl of concentrated virus) by qRT PCR.

Price for academic users: Basic fee: 62 € +4 €/ sample

Lentivirus replication-competent virus (RCV) test

RCV test is mandatory for transduced cells before removing them from BSL2 laboratory. RCV testing is based on qRT PCR in our Core. The customer should provide 10 to 100 µl of cell culture medium for testing.

Price for academic users: Basic fee: 62 € +1 €/ sample

Lentivirus production

While we primarily aim to guide and train our users to produce their own viruses, we also perform lentivirus production. The customer brings the transfer vector with the cloned-in gene of interest, and Genome Editing Core produces lentivirus particles using third third-generation packaging system. The users are responsible for getting permission for using genetically modified micro-organisms according to Finnish legislation.

Price for academic users:

• Lentivirus production, concentration (ultracentrifugation) + physical titer determination (qPCR): 450 € / virus. The yield is around 10^6 – 10^7 virus particles/ µl, and our customer receives 10 tubes of 10 µl virus aliquots.
• Lentivirus production (supernatant) + physical titer determination (qPCR): 300 €/ virus. The yield is around 10^7 – 10^8 virus particles/ ml, and our customer receives 10 tubes of 1.5 ml virus aliquots.
• Lentivirus production (supernatant) only: 150 €/ virus. The yield is undetermined, and our customer receives 10 tubes of 1.5 ml virus aliquots.

CRISPR/Cas9 genome editing

CRISPR knock-out

To knock out a gene-of-interest using the CRISPR/Cas9 technology, Cas9 nuclease is introduced into cells, creating double-strand breaks to a specific site in the genomic DNA guided by a guide RNA (sgRNA). Incorrect repair of these double-strand breaks generates small indels in the genomic DNA, leading to premature stop codons and loss of mature protein. We offer services in non-viral gene editing method, where the Cas9 ribonucleoprotein complex (RNP; Cas9 + guideRNA) is assembled in vitro, and is then introduced directly to the cells using nucleofection. This method offers fast genome editing by delivering a functional nuclease that degrades quickly, reducing the possibility of off-target effects. We help our customers with sgRNA sequence design, experimental implementation, and result analysis. The general reagents (e.g. Cas9 enzyme, tracrRNA, negative control crRNA, etc.) can be purchased according to the need.

The price for academic users will be in the range of 400 – 500 € depending on how much of the practical work is done by the core staff.

CRISPR knock-in

CRISPR knock-in, such as mutations into endogenous genes or endogenous epitope tagging, is achieved based on in vitro Cas9 ribonucleoprotein (RNP) assembly and delivery (nucleofection) to target cells. Here a single-stranded DNA (ssDNA) repair template of 100-200 bases, containing the desired edits to the genomic locus, is also introduced to the cells along with the Cas9 RNP. DNA repair machinery uses the introduced ssDNA as a template to repair the double-strand breaks created by the Cas9, and the desired edits are incorporated into the genomic DNA. We help our customers with sgRNA and target template sequence design, experimental implementation, and result analysis. The general reagents (e.g. Cas9 enzyme, tracrRNA, negative control crRNA, etc.) can be purchased according to the need.

The price for academic users is in the range of 500 – 700 € depending on how much of the practical work is done by the core staff.

Nucleofection

Nucleofection is an electroporation technique for introducing biological material to hard-to-transfect cells. The core has a Lonza 4D-nucleofector  instrument which has a variety of predefined programs for the electroporation of wide range of different cell types in a small 20 µl volume. The core offers aliquots of nucleofection reagents for testing or for experiments.

Price for academic users:             16 € / reaction in 20 µl volume

Training

The Genome Editing Core has wide experience in working with lentiviruses, retroviruses, and adeno-associated viruses. Additionally, our expertise includes CRISPR/Cas9 genome editing with non-viral methods. We provide training on viral production or CRISPR/Cas9 genome editing methods as a service to customers.

Price for academic users:              30 €/ hour