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. 2016 Aug 3;113(33):E4828–E4836. doi: 10.1073/pnas.1609792113

Fig. 2.

Fig. 2.

Genomic disruption of miR-309 by CRISPR/Cas9 blocked ovarian development in mosquitoes. (A) The graphical representation of scaffold of miR309-specific sgRNA. Tss indicated the transcriptional start site at the 5′-terminal end to facilitate in vitro transcription by T7 RNA polymerase. The sequence in green is the base-pairing region of sgRNA that targets miR-309 mature sequence, which is close to protospacer-adjacent motifs (PAMs) of “NGG” nearby. (B) Sequence alignment of sgRNA-targeted genomic region. (C and D) Ovaries were dissected from WT female mosquitoes at 36 h PBM and 72 h PE. (E) Ovaries were dissected from miR-309 mutant female mosquitoes. MG, midgut; Ov, ovary. (F and G) Alexa Fluor 488-labeled phalloidin staining ovaries from WT at 72 h PE and miR-309 mutant at 36 h PBM. The fluorescence imaging was visualized under the Zeiss microscope. (Scale bar: 0.5 mm.)