Compositions and methods for modulating mecp2 expression
a composition and mecp2 technology, applied in the field of oligonucleotide based compositions, can solve the problems of slowed brain and head growth, seizures, ambulatory problems, etc., and achieve the effect of relieving or enhancing the expression of mecp2 and preventing the mediated repression of mecp2
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[0193]The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.
Materials and Methods:
Real Time PCR
[0194]RNA is harvested from the cells using Promega SV 96 Total RNA Isolation system or Trizol omitting the DNAse step. RNA harvested from cells is normalized so that equal RNA is input to each reverse transcription reaction. Reverse transcriptase reactions are performed using the Superscript II kit and real time PCR performed on cDNA samples using icycler SYBR green chemistry (Biorad). A baseline level of mRNA expression for MECP2 is determined through quantitative PCR as outlined above. Baseline levels are also determined for mRNA of various housekeeping genes which are constitutively expressed. A “control” housekeeping gene with approximately the same level of baseline expression as the target gene is chosen for comparison purposes.
Cell Culture
[0195]Human hepatocyte Hep3B, human hepatocyte HepG2 cells, mouse h...
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