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RNA interference of MRCK‐1 in Caenorhabditis elegans to explore oxidative stress responses (802.26)
Author(s) -
Robben Michael,
Erickson Patti
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.802.26
Subject(s) - caenorhabditis elegans , green fluorescent protein , rna interference , microbiology and biotechnology , oxidative stress , biology , heat shock protein , rac1 , gene knockdown , small hairpin rna , reporter gene , biochemistry , signal transduction , gene expression , rna , gene
The Caenorhabditis elegans myotonic dystrophy kinase‐related, Cdc42 binding kinase (MRCK‐1) is activated by the Rho GTPase CDC‐42 (cell division control protein 42), which contains a conserved redox‐sensitive sequence motif on the phosphoryl binding loop. If reactive oxygen species (ROS) can activate CDC‐42, MRCK‐1 may be required to elicit a downstream response to oxidative stress. To test this, mrck‐1 expression was targeted using two RNA interference (RNAi) constructs in transgenic lines of C. elegans that produce green fluorescent protein (GFP) driven by redox‐sensitive promoters. After heat‐shock and chemically‐induced oxidative stress, GFP expression levels were quantified using epifluorescent microscopy. Preliminary data from heat‐shock protein reporter strain CL2070 ( hsp‐16.2 :GFP) indicate no significant GFP expression changes in the RNAi‐treated worms before or after heat stress, but further studies with different oxidative stressors and reporter strains are underway. If reduction of mrck‐1 is found to affect oxidative stress responses, a new mechanism for prevention of degenerative diseases and aging could be suggested. Grant Funding Source : Supported by the Henson Undergraduate Research fund.