Name | Variable | 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 | |||
NB_MOLECULES | 5,000,000 | >0 | number of expressed RNA molecules simulated | |
EXPRESSION_K | -0.6 | exponent of the expression power law ("Pareto coefficient") | ||
EXPRESSION_X0 | 9,500 | parameter of the exponential decay | ||
EXPRESSION_X1 | 9,5002 | parameter of the exponential decay | ||
TSS_MEAN | ||||
POLYA_SCALE | ||||
POLYA_SHAPE |
Input: reference annotation (REF_FILE), transcript filtering parameter (LOAD_CODING, LOAD_NONCODING), expression parameters (NB_MOLECULES, EXPRESSION_K, EXPRESSION_X0, EXPRESSION_X1)
In the beginning, the Flux Simulator reads the transcripts of the reference annotation (REF_FILE) and clusters genomic overlapping ones into loci. Transcripts that are annotated as non-/coding can be selectively disregarded (LOAD_CODING, LOAD_NONCODING). Then to assign a random expression profile where not necessarily all transcripts of the reference are expressed. Expression levels are connected with the relative expression rank by a mixed power- and exponential law of the general form
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.
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.
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).
Output: One line per simulated transcript molecule containing in the LIB_FILE.