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Fig. 7
Morpholino targeting of miRNA maturation in zebra fi sh. (
a
) Schematic of
pri-miRNA with different morpholinos targeting different portions of the sequence,
affecting both Drosha and Dicer processing. (
b
) Effect of the different morpholi-
nos on the miR-205 levels assessed by northern blot analysis of 30-h-old
embryos. (
c
) EGFP assay of embryos injected with morpholino and GFP-pri-
miR-205 construct. The miRNA activity is decreased upon morpholino co-
injection, resulting in an increased EGFP signal. Image adapted with permission
from Kloosterman WP et al. (2007) PLoS Biol
the morpholinos directly targeting the mature miR-375 only slightly
decreased the expression of miR-375; however the morpholino was
able to inhibit the function of miR-375 in a GFP reporter assay.
The knockdown of various miRNAs in zebra fi sh embryos by
morpholinos did not result in gross morphological deformities,
showing no signifi cant effect on embryonic development after
4 days. While there were initially no developmental defects 5 days
after morpholino-mediated knockdown of miR-375, 7 days after
injection a majority of the embryos died. Further investigation
