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Parameter

Parameter

Name

Default

Value

Parameter

Range

Description
REF_FILE
  file from which the reference annotation (GTF format) is read
LOAD_CODING
true{true,false}flag to dis-/consider transcripts that have an annotated coding sequence
LOAD_NONCODING
true{true,false}flag to dis-/consider transcripts that are annotated to be non-coding
PRO_FILE
  file to which the simulated expression values are written
LIB_FILE
  file to which the expressed transcript molecules are written

The Distribution of Gene Expression Levels

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Parameter

Name

...

Default

Value

...

Parameter

Range

...

NB_MOLECULES

...

EXPRESSION_K

...

EXPRESSION_X0

...

EXPRESSION_X1

...

Section

Input: reference annotation (REF_FILE), transcript filtering parameter (LOAD_CODING, LOAD_NONCODING), expression parameters (NB_MOLECULES, EXPRESSION_K, EXPRESSION_X0, EXPRESSION_X1)

...

Overview

Section

The "Gene Expression" step employs the annotation specified by REF_FILE_NAME and creates an artificial expression profile of the described transcripts. By the flag LOAD_CODING transcripts with an annotated coding sequence are taken into account, by LOAD_NONCODING correspondingly those which don't. Results from in silico gene expression are stored in the files specified by the parameters PRO_FILE respectively LIB_FILE; if no explicit values are provided for these parameters, then the corresponding files are created in the folder of the parameter file.

Details

Section

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Section

where denotes the rank number of a gene and is the exponent of the intrinsic power law, and respectively  control the exponential decay. The Flux Simulator assigns to the transcripts in the reference annotation randomly expression ranks which then are turned into relative expression levels by the modified Zipf's Law above, which determines the initial number of molecules by multiplication with the total numbers of molecules. Default values for parameters and have been estimated for mammalian cells by non-linear fitting to expression levels observed in experimental results.

Section

Output: Columnn 1-6 of the PRO_FILE, i.e., (1) locus name, (2) transcript identifier, (3) coding flag, (4) length of the processed transcript, (5) relative fraction  and (6) absolute number of the transcript species in the initial RNA extraction.

Transcript Modifications during Expression

Parameter

Name

Default

Value

Parameter

Range

Description
TSS_MEAN
25>0rate of the exponential for deviation of simulated transcription starts from annotated transcription start point
POLYA_SCALE
300>0scale parameter of the Weibull distribution describing poly-A tail lengths
POLYA_SHAPE
2>0shape paramter of the Weibull distribution describing poly-A tail lengths
Section

Input: Columnn 6 of the PRO_FILE, i.e., the absolute number of RNA molecules that is simulated for a certain transcript in the experiment and the parameters of transcription start (TSS_MEAN) and poly-A tail variation (POLYA_SCALE, POLYA_SHAPE).

Section
Based on the number of RNA copies that is simulated for each transcript, genes are in silico expressed. In this process,  assigned individual variations in transcription start and the length of the attached poly-A tail. The Flux Simulator models differences in the annotated transcription starts by an exponential distribution with an adjustable mean value (TSS_MEAN). During poly-adenylation in the nucleus usually 200-250 adenine residues get added to the primary transcript. Disregarding other poly-adenylation mechanisms (e.g., cytoplasmatic polyadenylation) the Flux Simulator describes poly-A lengths by a flexible Weibull distribution (POLYA_SCALE, POLYA_SHAPE).
Section

Output: One line per simulated transcript molecule containing in the LIB_FILE.