SERUM AMYLOID P COMPONENT (APCS) iRNA COMPOSITIONS AND METHODS OF USE THEREOF
a technology of amyloid p component and irna composition, which is applied in the direction of peptides, biochemistry apparatus and processes, organic active ingredients, etc., can solve the problems of affecting the function of tissues and organs, no amyloidosis cure, and often delayed diagnosis
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iRNA Synthesis
Source of Reagents
[0528]Where the source of a reagent is not specifically given herein, such reagent can be obtained from any supplier of reagents for molecular biology at a quality / purity standard for application in molecular biology.
[0529]A set of siRNAs targeting the human SAP or APCS, “amyloid P component, serum” (human: NCBI refseqID NM_001639; NCBI GeneID: 325) as well as toxicology-species APCS orthologs (cynomolgus monkey: XM_005541312; mouse: NM_011318; rat: NM_017170) were designed using custom R and Python scripts. The human NM_001639 REFSEQ mRNA, version 3, has a length of 960 bases. The rationale and method for the set of siRNA designs is as follows: the predicted efficacy for every potential 19mer siRNA from position 10 through the end was determined with a linear model derived the direct measure of mRNA knockdown from more than 20,000 distinct siRNA designs targeting a large number of vertebrate genes. A subset of the APCS siRNAs was design...
example 2
iRNA Design, Synthesis, Selection, and In Vitro Evaluation
[0534]An additional set of siRNAs targeting the human APCS (SAP), “amyloid P component, serum” (human: NCBI refseqID NM_001639.3; NCBI GeneID: 325) as well the toxicology-species APCS ortholog from cynomolgus monkey: XM_005541312.2) was designed using custom R and Python scripts. All the siRNA designs are a perfect match to the human APCS transcript and a subset have either perfect or near-perfect matches to the cynomolgus monkey ortholog. The human NM_001639 REFSEQ mRNA, version 3, has a length of 960 bases.
[0535]A detailed list of the unmodified SAP sense and antisense strand sequences is shown in Table 6. A detailed list of the modified SAP sense and antisense strand sequences is shown in Table 7.
In Vitro Screen
[0536]Dual-Glo® Luciferase Assay
[0537]Cos7 cells (ATCC, Manassas, Va.) were grown to near confluence at 37° C. in an atmosphere of 5% CO2 in DMEM (ATCC) supplemented with 10% FBS, before being released...
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