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Project:

Background:

  • AGO4 is the effector in Pol IV-dependent sRNAs mediated RdDM pathway;
  • Pol IV, RDR2, and DCL3 sequentially contribute to the production of Pol IV-dependent sRNAs, which, in recent model, can stabilize the AGO4 protein in cell;
  • pol iv and rdr2 mutants eliminate AGO4, while DCL3 only partially decreases the AGO4, therefore other sRNAs might be produced and stabilize AGO4;
  • DCL3 homologous DCL2 and DCL3 also produce different kinds of sRNAs.

Aim:

  • to explore the potential sRNAs loaded  to AGO4 in dcl3 mutant, as these sRNAs might contribute to the stabilization of AGO4 protein in cell.
  • Problem to solve: how do the profiles of sRNAs (as well as their origin locus) in dcl3 and dcl2/3/4 mutants change?

Methods:

  • by compare the sRNAs groups based on length or gene/sequence types between dcl3 and dcl2/3/4 mutants, we will find out the sRNAs deduced in dcl2/3/4, which might load to AGO4 and contribute the stabilization of AGO4 in dcl3 mutant.
  • Note: we unfortunately can't find out the data for sRNAs loaded to AGO4 in dcl3 and dcl2/3/4 mutants, which will directly indicate the change of sRNAs loaded to AGO4, giving that AGO4 is stabilized when sRNAs are loaded.

Concept map:

Reference:

1, Groth M. et al, SNF2 chromatin remodeler-family proteins FRG1 and -2 are required for RNA-directed DNA methylation. PNAS. 2014. 111(49): 17666-71.