About mos1-mediated Single Copy Insertion (mosSCI)
mosSCI is a method to insert a single copy of a transgene into a well-defined location in the C. elegans genome. The method has the advantages:
- Transgenes can be expressed at levels close to endogenous gene expression
- Transgene insertions are stable
- Transgene expression is possible in the germline.
The method works by breaking a chromosome at a particular location by excising a Mos1 transposon. The excision creates a double-strand DNA break that is repaired by the cell. In the presence of a DNA template with homology to the breakpoint then the repair process will incorporate DNA from the repair template into the genome. We have incorporated a positive selection marker to select for this event (typically unc-119) and fluorescent markers to identify extra-chromosomal arrays.
Requirements
- A transgene cloned into the appropriate targeting vector.
- A strain containing a Mos1 transposon at the site corresponding to the targeting vector.
Schematic of method
