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Product Information
Product Usage Information
CST recommends transfection with 100 nM SignalSilence® DNMT3L siRNA I (Mouse Specific) 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.
Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.
Storage
Specificity / Sensitivity
Species Reactivity:
Mouse
Product Description
Background
Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting, and development (1,2). Three families of mammalian DNA methyltransferases have been identified: DNMT1, DNMT2, and DNMT3 (1,2). DNMT1 is constitutively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methylation patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic stem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as de novo methyltransferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither de novo nor maintenance DNA methylation.
DNMT3L is a catalytically inactive regulatory factor for the DNMT3A and DNMT3B de novo methyltransferases that is expressed at low levels in embryonic stem cells, testis, ovaries, and thymus (1,2). These de novo methyltransferases consist of a heterotetrameric complex containing two molecules of DNMT3L, and either two molecules of DNMT3A or DNMT3B (3). DNMT3L contains an amino-terminal ATRX-DNMT3-DNMT3L (ADD) domain and a carboxy-terminal methyltransferase-like domain (4-7). The methyltransferase-like domain binds to DNMT3A and DNMT3B to stimulate catalytic activity by increasing the binding of S-adenosylmethionine and DNA (4,5). The ADD domain recruits the methyltransferase complex to transcriptionally inactive regions of the genome by binding to unmethylated histone H3 Lys4 (6,7).
- Hermann, A. et al. (2004) Cell Mol Life Sci 61, 2571-87.
- Turek-Plewa, J. and Jagodziński, P.P. (2005) Cell Mol Biol Lett 10, 631-47.
- Jia, D. et al. (2007) Nature 449, 248-51.
- Holz-Schietinger, C. and Reich, N.O. (2010) J Biol Chem 285, 29091-100.
- Suetake, I. et al. (2004) J Biol Chem 279, 27816-23.
- Ooi, S.K. et al. (2007) Nature 448, 714-7.
- Otani, J. et al. (2009) EMBO Rep 10, 1235-41.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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