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Jump-out of the P-element was indicated by loss of white eye marker.
In the second step, the eye marker was replaced with DNA sequences of choice using RMCE enabling flexible gene modification.
In the F2 progeny, flies expressing GFP, RFP fluorescent markers, and the red eye marker were the conditional mutant flies.
In addition to precise RMCE of the markers, a third line (Dm-loxM1b) showed only loss of the eye marker.
We also confirmed that depletion of xWNK4 reduced expression of neural marker genes such as pan-neural marker, NCAM; forebrain marker, BF-1; eye marker, Rx1 (Fig. 3F).
To obtain transgenic gRNA flies, the five pUAST-U6B/CR7T-gRNA vectors were separately injected into w fly embryos, and at least three independent lines per transgenic gRNA were identified through observation of the RFP eye marker.
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Fixed eye markers of the face model are used if an eye could not be detected by the eye search.
The introduction of gel liners, felt-tip eye markers and angled brushes like Kashuk's Longwear Gel Liner and Bent Eyeliner Brush has made the cat-eye much easier to do today.
The female red-eye vasa -Cas9 flies were crossed to pUAST-CR7T-gRNA males to generate male F0 flies, which were identified by screening for the GFP and RFP eye markers.
As with the morpholino injections, misexpression of sol-efnb2a led to delayed, asymmetric optic vesicle evagination and to the presence of cells expressing eye markers within the brain (Fig. 2D).
A survey of spatiotemporal distributions of Vmem during craniofacial development revealed two spots of hyperpolarized cells (lighter stain) that become eyes; artificial depolarization of these cells by misexpression of a cation channel altered expression of endogenous eye markers and induced eye defects.
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