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Transformation of Nematodes With the Helios Gene Gun

While the majority (up to 90%) of C. elegans hermaphrodites are killed during Biolistic DNA transfer, the survivors with their gold particles incorporated may be cultivated on new E. coli-seeded NGM agar plates (Figure 2A shows particles within a living hermaphrodite). F1/F2 progeny nematodes were screened 3 to 4 days later for visible dominant mutant marker rol-6 (Kramer et al., 1990 ). Animals carrying roller allele su-1006 exhibit a helically twisted cuticle and body. Muscle contractions, which would normally propel the worm forward or backward in a sweeping sinusoidal motion, will cause the nematode instead to roll over its longitudinal axis and to move in circles (Mello et al., 1991; Fire, 1986; Kramer et al., 1990). In the case of L. sigmodontis, no special selection mode is necessary because the success of bombardment can be visualized by standard light microscopy to make visible the gold particles now located within these filariae (see Figure 2B).


Results
The established mode of DNA transfer into C. elegans is microinjection of DNA into the gonadal syncytium (Stinchcomb et al., 1985; Fire, 1986). Under standard selection conditions promoter-reporter gene cDNA constructs are used, in co-transfection with a visible dominant such as mutation rol-6 (su1006), which leads to roller movement phenotype of transformed worms (already in F1 generation) and is easily detectable within a background of normally moving animals. The same transfection efficiency has been observed for this Biolistic DNA transfer method as for microinjection experiments (Fire, 1986), while this new strategy is less time consuming and much more convenient for co-transfection experiments. In the F1 generation,
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