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Discover the pathway involving DICER and RISC, explore software for siRNA design, databases, and methods for siRNA delivery and testing. Design and test siRNA to suppress ZF efficiently.
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Recovery of ZF by PCR FLAG+ FLAG-
RNA interference May 9
siRNA • discovery • pathway DICER and RISC • software for siRNA design • databases • delivery of siRNA • testing siRNA • designing and testing siRNA against ZF
suppression of ZF by siZF 10ng 100ng 500ng pcZF psiZF#14 µg 0 0.5 1 0 0.5 1 0 1 0.5 1 0.5 1 psiRad9 µg 1 2 3 4 5 6 7 8 9 10 11 14
U6 prom GFP mU6pro BbsI XbaI PolIII term BspI TTTG…………TTTT …………AAAAGATC XbaI si hairpin
siRNA for ZF ATGGAGGAGGAGGCGATCGCCAGCCTCCCGGGGGAAGAGACGGAGGATATGGACTTTCTGTCTGGGCTGGAACTGGCGGATCTCCTGGACCCCAGGCAACCGGACTGGCACCTGGACCCCGGGCTTAGCTCGCCGGGGCCTCTCTCCTCGTCTGGCGGAGGCTCGGATAGCGGCGGCCTGTGGAGAGGGGACGATGACGATGAGGCCGCGGCTGCTGAAATGCAGCGCTTCTCTGACCTGCTGCAAAGGCTGTTAAACGGTATCGGAGGCTGCAGCAGCAGCAGTGACAGTGGCAGCGCCGAAAAGAGGCGGAGAAAGTCCCCAGGAGGAGGCGGCGGTGGCGGCAGCGGTAACGACAACAACCAGGCGGCGACAAAGAGTCCCCGGAAGGCGGCGGCGGCCGCTGCCCGCCTTAATCGACTGAAGAAGAAGGAGTACGTGATGGGGCTGGAGAGTCGAGTCCGGGGTCTGGCAGCCGAGAACCAGGAGCTGCGGGCCGAGAATCGGGAGCTGGGCAAACGCGTACAGGCACTGCAGGAGGAGAGTCGCTACCTACGGGCAGTCTTAGCCAACGAGACTGGACTGGCTCGCTTGCTGAGCCGGCTGAGCGGCGTGGGACTGCGGCTGACCACCTCGCTCTTCAGAGACTCGCCCGCCGGTGACCACGACTACGCTCTGCCAGTGGGAAAGCAGAAGCAGGACCTGCTGGAAGAGGACGACTCGGCGGGAGGAGTCTGTCTCCATGTGGACAAGGATAAGGTGTCGGTGGAGTTCTGCTCGGCGTGCGCCCGGAAGGCGTCGTCTTCTCTTAAAATGTAG #14 #15