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. 2014 Apr 7;7:25. doi: 10.3389/fnmol.2014.00025

FIGURE 4.

FIGURE 4

In vitro screening strategy to identify a potent shRNA knockdown sequence. (A), a construct expressing the GCAP1-EGFP fusion protein under the control of the CMV promoter. (B), the shRNA expression construct. shRNA (hp yellow box) is under the control of the H1 promoter, and the reporter gene mCherry is under the control of the CMV promoter. Hp consists of a sense-loop-antisense construct (the hairpin). Tts, transcription terminator signal. (C), four candidate shRNAs for suppression of bovine GCAP1, and a 2-nucleotide mismatch control. (D), four candidate shRNAs for suppression of both wild-type and mutant mouse Guca1a mRNA, and a 2-nucleotide mismatch control. See also Figure 1 (Jiang et al., 2011), and Figure 6 (Jiang et al., 2013).