This motif discovery algorithm selects the final motif with most distinguished feature (total accessibility or site number) in bound (positive) and unbound (negative) transcripts.
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Sequence accessibility of transcripts is predicted by the RNAplfold tool in the ViennaRNA package.
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pos_file: A file that contains gene names in the positive set, one gene per line.
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neg_file: A file that contains gene names in the negative set, one gene per line.
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seq_file: A fasta file containing sequences of transcripts in positive and negative sets (in the RNA alphabet). Make sure gene names in this file are consistent with those in pos_file and neg_file.
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RNAplfold_direct: A folder containing the
<ID>_lunp
files of RNAplfold output of genes in positive and negative sets. Make sure gene names in this file are consistent with those in pos_file and neg_file.To get result from RNAplfold, assuming the length of the binding site is at most 10 nt:
RNAplfold -W 80 -L 40 -u 10 < seq_file_name
The setting of parameters W, L, u can be changed upon different situation.
For detailed information of running RNAplfold, please refer to https://www.tbi.univie.ac.at/RNA/RNAplfold.1.html.
To run the code, you will need to specify the following parameters by modifying the following lines of run2_new_RNAplfold_format.py
:
model = 'access_seq'
pos_file_name = '' # name of pos_file
neg_file_name = '' # name of neg_file
seq_file_name = '' # name of seq_file
RNAplfold_direct = '' # directory name of the RNAplfold results
final_out_name = '' # name of the final output (only the best motif)
detailed_final_out_name = '' #name of the detailed output (motifs from all five seed 6mers)
To run the code, one need to run the following function in the run2_new_RNAplfold_format.py
,
Motif_discovery(final_out_name, detailed_final_out_name, model, pos_file_name, neg_file_name, seq_file_name, RNAplfold_direct)
X. Li, G. Quon, H.D. Lipshitz, Q.D. Morris, Predicting in vivo binding sites of RNA-binding proteins using mRNA secondary structure, RNA 16.6 (2010): 1096–1107. [Pubmed] [Supplementary materials]