| 主权项 |
1. A method of evaluating an effect of a test compound on an endogenous RNA subset associated with an mRNA-protein (mRNP) complex in a cell comprising an endogenous RNA binding protein (RBP), endogenous mRNP complex associated protein, or an endogenous RNA associated protein (RAP), the method comprising the steps of:
a) exposing the cell to the test compound; b) lysing the cell exposed to the test compound to produce a lysate; c) contacting the lysate with a ligand that specifically binds at least one component of the mRNP complex, wherein the mRNP complex is bound by the ligand; d) partitioning the mRNP complex bound by the ligand to a solid support; e) removing the mRNP complex bound to the ligand and the solid support from the lysate; f) identifying and, optionally quantifying, the endogenous RNA subset from the removed mRNP complex without amplifying the RNAs by PCR, wherein the identified RNAs of the endogenous RNA subset are encoded by a plurality of distinct genes; and g) comparing the identified RNAs of step f) with the RNAs from a control RNA subset, and optionally the levels of one or more RNAs in step f) to the levels of the one or more RNAs in the control RNA subset, from the mRNP complex generated by performing steps b-f on a control cell, wherein the control cell is of the same type as the test cell and is not exposed to the test compound; thereby evaluating the effect of the test compound on the identity, and optionally the quantity, of the endogenous RNA subset. |
