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Watch our archived webinar on genome editing in hard-to-transfect cells:
Duration: 61 minutes
The CRISPR (clustered regularly interspaced palindromic repeats) technology is based on a bacterial defense pathway against viral invasion. It has been adapted for genome editing applications in eukaryotic cells and is an emerging technology because it allows for some more flexibility compared to zinc finger nucleases (ZFNs) or transcriptional activator-like effector nucleases (TALENs).
In contrast to ZFN and TALEN, CRISPR-based genome editing is relying on two separate components working together:
Since the DNA-specific part is an RNA molecule it is much easier to design and produce than a ZF- or TALE-nuclease fusion protein. In addition, by transferring Cas9 nuclease together with several gRNAs targeting different sites multiple genome editing is possible.
CRISPR-based Genome Editing Requires Co-transfection
For CRISPR-based genome editing, various transfection scenarios are possible:
Our Nucleofector™ Technology has been shown to work as a reliable transfection method for CRISPR-based genome editing tools
The Nature Protocols publication from Ran et al.3 provides a very comprehensive guideline on using the 4D-Nucleofector™ System in combination with CRISPR technology . For further technical assistance on using the Nucleofector™ Technology for CRISPR-based genome editing, contact Lonza Scientific Support.
1) Petit CS et al. (2013) J Cell Biol 202:1107-1122 (cells: HeLa)
2) Ran FA et al. (2013) Cell 154:1380–1389 (cells: various cell lines, e.g. HEK293FT)
3) Ran FA et al. (2013) Nat Prot 8(11):2281–2308 (cells: HEK293 and HUES62)