Fig. 3: Germline expression of I-AniI induces substantially more SSA and NHEJ than I-SceI in the P5-EGFP transgenic locus. | Communications Biology

Fig. 3: Germline expression of I-AniI induces substantially more SSA and NHEJ than I-SceI in the P5-EGFP transgenic locus.

From: Transgene removal using an in cis programmed homing endonuclease via single-strand annealing in the mosquito Aedes aegypti

Fig. 3

a Schematic representation of the SSA-capable P5-EGFP transgene integrated into the Ae. aegypti genome containing two marker genes (eye-specific DsRED and whole body-expressed EGFP) interspersed with two groups of homing endonuclease target sites (site-A and site-B) were engineered to flank the EGFP cargo. b Percentage of F2 larvae with phenotypic loss of the EGFP marker from triplicate experiments. The X-axis indicates the grandparent (F0)/parent (F1) contributing the nuclease gene. Tukey’s multiple comparison test was found to be significant (2-way ANOVA, P < 0.001), and statistically different groups are marked (a and b). c HRMA of the site-A for F2 mosquitoes scored as WGR (white and DsRED+ eyes; EGFP+ body) by using a primer pair of SV40-F and PUb-5R. d Sequence analysis of amplicons with altered melting curves from (c), compared to the baseline. Sequencing analysis revealed various numbers of nucleotide deletions occurred at site-A. The colors of IDs designate individual HRMA peaks shown in (c).

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