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MicroRNA-Directed Cleavage of HOXB8 mRNA

MicroRNA-Directed Cleavage of HOXB8 mRNA. Soraya Yekta,I-hung Shih, David P.Bartel 23 APRIL 2004 VOL 304 SCIENCE. Hox genes in mammals. The vertebrate homeotic complex comprises four distinct Hox gene clusters (Hox A, B, C, D) that are organized into thirteen homology

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MicroRNA-Directed Cleavage of HOXB8 mRNA

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  1. MicroRNA-Directed Cleavage of HOXB8 mRNA Soraya Yekta,I-hung Shih, David P.Bartel 23 APRIL 2004 VOL 304 SCIENCE

  2. Hox genes in mammals • The vertebrate homeotic complex comprises four distinct Hox gene clusters (Hox A, B, C, D) that are organized into thirteen homology • The chromosomal organization of the genes in each Hox cluster reflects its anterior-posterior expression in the body plan (spatial colinearity). • In general, members of the same subgroup are expressed at the same time and have the same anterior boundary of expression

  3. The current model for the biogenesis and post-transcriptional suppression of microRNAs and siRNAs

  4. Recent progress • In plant, most miRNAs anneal to their targets with nearly complete complementarity at a single site,leading to mRNAs cleavage and degradation • The extent of complementarity between the siRNA/miRNA and its target can determine the mechanism of silencing

  5. miR-196 in Vertebrate

  6. The sequence and genomic context of the vertebrate miR-196 family Bold: residues of the mature miRNA; Red: miRNAs validated by cloning. Asterisks: perfectly conserved residues

  7. Overview of the 5’ RACE Procedure

  8. RNAse protection assay

  9. Expression of miR-196 during mouse embryonic development. The Northern blot probes total RNA from 7-day, 11-day, 15-day, and 17-day mouse embryos (lanes 1–4, respectively) M :33P-labeled RNA size markers

  10. MiR-196–directed cleavage of HOXB8 mRNA(1) blackAbsolutely conserved nucleotides; redcomplementary site ; pinkG:U wobble

  11. MiR-196–directed cleavage of HOXB8 mRNA(2) perfect antisense sequence of miR-196a: miR-196a-as perfect antisense sequence of miR-1d: miR-1d-as miR-1d expressed in skeletal muscle cells

  12. MiR-196–directed cleavage of HOXB8 mRNA(3)

  13. Additional HOX genes predicted to be regulatory targets of miR-196(1) Wide red bars extensive complementarity, narrow bars seed matches, Asterisks slightly relaxed seed matches that permit G:U wobble

  14. Sequence alignment of the miR-196 complementary site, and a representative complementary seed and flanking regions in the 3´ UTRs of HOXC8 and HOXD8 genes. Black:absolutely conserved nucleotides Red: miR-196 complementary site and the seed Pink:G:U wobbles

  15. Additional HOX genes predicted to be regulatory targets of miR-196(2)

  16. Additional HOX genes predicted to be regulatory targets of miR-196(3)

  17. Discussion • Transcriptional repression may also involved • Genetic model of mRNA cleavage in vivo • The difference of miRNA and siRNA • The mechanism of miRNA function

  18. Thank you!

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