Figure 1. Bashing and identification of a cis-regulatory region responsible for the rectal cell-specific
ceh-6 expression and rescue of
ceh-6 mutants transdifferentiation defects: (A) Schematic view of the
ceh-6 genomic fragment used for dissection of rectal cell-specific regulatory sequences. The in-frame insertion site of GFP, as well as the location of
ceh-6 ATG and TAG, and each region size are indicated. Deleted regions are represented as closed brackets containing their name in blue (e.g. 'a', 'b', 'c', or the names of the deleted intron
i1 to
i5 (intron 1-5) and 3'UTR deletion). The location of the deletions with respect to the ATG is also indicated under brackets on the right. Orange: the
ceh-6 3'UTR was swapped with
unc-54 3'UTR. (B) The ability of the various
ceh-6 constructs to i) rescue
ceh-6(
gk665) lethality; ii) drive expression in rectal cells and iii) rescue
ceh-6(
gk665) defects in transdifferentiation of the Y rectal cell into a PDA neuron is indicated. (n), total number of L3 animals and older scored. Note, constructs delta3'UTR led to transgenic lines that did not show any expression nor rescue of the lethality, and no lines could be obtained with construct
i1+
i4. (C) Expression pattern of
ceh-6 constructs in the Y and rectal cells. (i), expression pattern of syIs63 (
cog-1::gfp) alone for reference; (ii) WT control construct; (iii) 'c' construct; (iv), schematic representation of the rectal cells; Red arrowhead, Y cell. Dotted white line, rectal slit. All images were acquired at the L1 larvae stage. Scale bar represents 10 um.