Figure 1.
Cre-mediated floxed GFP inactivation.
(A) Pges-1::Cre was injected into tmIs942[floxed Pdpy-30::NLS::GFP::LacZ SCI] along with the Pges-1::DsRed marker. The arrowheads indicate the nuclei of the intestinal cells. The nucleic GFP signals were eliminated in the DsRed-labeled intestinal cells. The arrows indicate the nuclei of the body wall muscle in which the GFP signals were observed. (B) Pscm::Cre was injected into tmIs942[floxed Pdpy-30::NLS::GFP::LacZ SCI] along with the Pscm::mCherry marker. The arrowheads indicate the nuclei of the seam cells. The nucleic GFP signals were selectively eliminated in the seam cells expressing mCherry. Scale bars = 20 µm. (C) DNA excision was examined by PCR using primers spanning the floxed region. The PCR produced 200 bp-band when the floxed region was excised. Bands of 200 bp were specifically observed in Ex animals harboring Pges-1::Cre (C) and Pscm::Cre (D), but not in the wild-type N2 or tmIs942 parental strain. Six Ex(+) animals were analyzed for each strain. (E, F) The NLS::GFP signals were diminished in the intestine (E) and the hypodermis (F) depending on the heat-shock treatment. The arrowheads indicate the nuclei of the intestinal cells (E) and hypodermal cells (F). Scale bars = 20 µm.
Table 1.
Cre recombinase expression vectors driven by tissue specific and heat-shock promoters and transgenic strains.
Figure 2.
Cre-mediated Venus::H2B expression.
(A) Schematic illustration of the Pdpy-30<mCherry<venus::H2B tester vector. Cell type-specific Cre expression vectors were co-injected with the tester vector and corresponding cell type ECFP markers. Venus::H2B is co-expressed with cytosolic ECFP in the cells where Cre/loxP recombination occurs. (B) Venus::H2B expression was detected with ECFP in pharyngeal cells of the Ex[Pdpy-30<mCherry<venus::H2B;Pmyo-2::Cre;Pmyo-2::ECFP] animals (B) and in ventral nerve cord neurons of the Ex[Pdpy-30<mCherry<venus::H2B;Punc-4::Cre;Punc-4::ECFP] animals (C). Scale bars = 20 µm.
Figure 3.
sid-1 was conditionally knocked out by Cre expression.
(A) Schematic overview of floxed sid-1 SCI by UV/TMP methods. The floxed sid-1 plasmid was co-injected with positive/negative selection marker plasmids into the recipient strain ben-1(tm234);sid-1(tm2700);vps-45(tm246) to establish parental Ex strains. The obtained Ex strain, ben-1(tm234);sid-1(tm2700);vps-45(tm246);tmEx3452 was treated with a TMP solution followed by UV irradiation. Single-low-copy integration strains were selected under the positive/negative selection condition. (B) The relative amounts of the sid-1, vps-45, and ben-1 genes were determined using quantitative PCR, and the levels of the genes (normalized to the ama-1 gene) are presented as ratios to the N2 control. The error bars represent the SE of three independent experiments. (C, D) Percentages of animals exhibiting the Bli phenotype in the bli-3 RNAi (C), and the twitch of body wall muscle in the unc-22 RNAi (D). The error bars represent the SE of three independent experiments. The statistical analysis was performed using t-test.