CRISPR/Cas9: Vector preparation and cell line edition
CRISPR/Cas9 is a powerful tool for genome editing. It requires the construction of a DNA vector that codes both for Cas9 protein and a target specific RNA called crRNA (also called single-guide RNA when combined with another sequence, tracrRNA) that must bind only where editing is desirable. In this experiment, the Expert will design and construct the DNA vector that should allow for the edition of the sequence in the target cell's DNA.
Materials to provide
- Accession number of genes of interest
- Cell lines to transfect
- Antibodies and conditions
Further details on the project may be requested.
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- Bacterial clones and preparation of the guide vector with insert
- Map of the vector
- Donor vector
- Cell line edition and shipping cell lines in frozen vials (3 x 1ml)
- Data for sorting enrichment
- Gene knock-out PCR verification if cell selection is performed
- Monoallelic verification
- Western blot data
- Removal of selection marker
- Study following the specifications validated with the Expert
Get started with your CRISPR/Cas9 cell line edition
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