Looper is a DNA tandem repeat identification tool. Tandem repeats are important genomic sequences which have functional and evolutionary significance.
Looper is scripted in C++.
Looper captures a mathematical property exhibited by binary number representations of DNA tandem repeats. In the binary number or the 2-bit format of DNA sequences, nucleotides are represented by 4 different combinations of 2 binary digits (bits). The binary number representations of DNA tandem repeats conform to a unique division rule. Looper scans the genome in windows, converts each window sequence to a binary number and checks if the number qualifies the division rule. The detailed explanation of the algorithm is below and an overview of the method is depicted in the figure below.
$ pip install looper_ssrThe help message and available options can be accessed using
$ looper_ssr -h which gives the following output
usage: looper_ssr [-h] -i <FILE> [-o <FILE>] [-m <INT>] [-M <INT>] [-l <INT>] [-a] [-g <FILE>]
[--anno-format <STR>] [--gene-key <STR>] [--up-promoter <INT>]
[--down-promoter <INT>]
Required arguments:
-i, --input <FILE> Input sequence file. Fasta format.
Optional arguments:
-o, --output <FILE> Output file name. Default: Input file name + _looper.tsv
-m, --min-motif-size <INT> Minimum size of a repeat motif in bp. Default: 1
-M, --max-motif-size <INT> Maximum size of a repeat motif in bp. Default: 6
-l, --min-length <INT> Cutoff repeat length. Default: 2*M.
Should at least be twice of maximum motif size.
-a, --analyse Generate a summary HTML report.
Compound repeat arguments:
--compound Report compound repeats. The output of compound repeats is a separate file with the suffix ".compound".
-d <INT>, --comp-dist <INT>
Maximum distance between individual repeats of compound repeat. Use negative to denote overlap. Default: 0
Annotation arguments:
-g, --annotate <FILE> Genomic feature file to annotate repeats w.r.t genes.
Both GFF and GTF can be processed.
--anno-format <STR> Format of genomic feature file.
Valid inputs: GFF, GTF. Default: GFF
--gene-key <STR> Attribute used as unique identifier for gene name.
The default identifier is "gene".
--up-promoter <INT> Upstream distance(bp) from TSS to be considered as
promoter region. Default: 1000
--down-promoter <INT> Downstream distance(bp) from TSS to be considered as
promoter region. Default: 1000
Expects: STRING (to be used as filename)
Default: None
This is the only required argument for the program. The input file must be a
valid FASTA file.
Expects: STRING (to be used as filename)
Default: Input Filename + _looper.tsv (see below)
If this option is not provided, the default output filename will be the same as the input filename, with its extension replaced with '_looper.tsv'. For example, if the input filename is my_seq.fa, the default output filename will be my_seq.fa_looper.tsv. If the input filename does not have any extension, _looper.tsv will be appended to the filename. Please note that even in the case of no identified SSRs, the output file is still created (therefore overwriting any previous file of the same name) but with no content in the file.
The output is a tab-delimited file, with one SSR record per line. The output columns follow the BED format. The details of the columns are given below:
| S.No | Column | Description |
|---|---|---|
| 1 | Chromosome | Chromosome or Sequence Name as specified by the first word in the FASTA header |
| 2 | Repeat Start | 0-based start position of SSR in the Chromosome |
| 3 | Repeat Stop | End position of SSR in the Chromosome |
| 4 | Repeat Class | Class of repeat as grouped by their cyclical variations |
| 5 | Repeat Length | Total length of identified repeat in nt |
| 6 | Repeat Strand | Strand of SSR based on their cyclical variation |
| 7 | Motif Number | Number of times the base motif is repeated |
| 8 | Actual Repeat | Starting sequence of the SSR irrespective of Repeat class and strand |
An example output showing some of the largest repeats from Drosophila melanogaster is given below
X 22012826 22014795 ACTGGG 1969 - 328 TCCCAG
2RHet 591337 591966 AATACT 629 - 104 ATTAGT
4 1042143 1042690 AAATAT 547 + 91 AAATAT
2RHet 598244 598789 AATACT 545 - 90 AGTATT
XHet 122 663 AGAT 541 + 135 GATA
X 22422335 22422827 AGAT 492 + 123 GATA
3R 975265 975710 AAAT 445 - 111 TTAT
X 15442288 15442724 ACAGAT 436 + 72 ACAGAT
2L 22086818 22087152 AATACT 334 - 55 TATTAG
YHet 137144 137466 AAGAC 322 - 64 CTTGT
Expects: INTEGER
Default: 1
Minimum length of motifs to be considered. By default, looper ignores redundant
motifs. For example, a stretch of 12 A's is considered a monomer repeat of 12
A's rather than a dimer repeat of 6 AA's.
Expects: INTEGER
Default: 6
Maximum length of motifs to be considered. Setting a large value of -M has a
non-trivial effect on both the runtime and memory usage of looper.
Expects: INTEGER
Default: 2 * M
Minimum length cut-off to be considered when finding an SSR. The same cut-off
will apply for SSRs of all motif lengths, even if the motif length is not a
divisor of this value. In such cases, SSRs that end with a partial motif are
also picked if they pass the length cut-off. This value should be at least twice
of the maximum motif size.
Expects: None
Default: False
In addition to the default tab-separated output, Looper can also generate a fully
interactive HTML report for easy downstream analysis of the repeat data. The
filename will be the same prefix as that of the main output. For example, if the
input filename was my_seq.fa, the analysis report will be my_seq_looper.html. An
example HTML report, generated from the repeat data of Homo sapiens (build hg19),
can be accessed here (Right click -> Save As).
Expects: None
Default: False
This is flag which when set to true reports all compound repeats. Compound repeats
are repeats which are either overlapping or separated by a small gap. Compound repeats
are reported in a separate file with the extension ".compound". The maximum distance
between two individual repeats of a compound repeat can be set using the option -d.
The compoud repeat has four columns with first four denoting the sequence, start,
end and length of the repeat. The fifth column denotes the repeat classes of individual
repeats of the compound repeat and the number of different cyclical variations that
have occured as compound repeat. The sixth column is the strand of the individual
repeats. The seventh column is denotes the actual motifs of individual repeats,
repeat length and the distance between the individual repeats. Below is an example
output reporting compound repeats.
chr1 94808 94847 (AAAAG)1(A)1 39 -|- (CTTTT)20|D0|(T)23
chr1 113871 113896 (AGAGGG)1(AAAGAG)1 25 -|- (CCCTCT)13|D0|(TCTCTT)12
chr1 130955 130977 (AAAAAC)1(AAAAC)1 22 +|+ (CAAAAA)12|D0|(AAAAC)13
chr1 147433 147475 (AAC)1(AAAAAC)1(AAC)1 42 +|+|+ (CAA)21|D0|(AACAAA)17|D-2|(AAC)14
chr1 156944 156967 (AAAAAT)2 23 +|+ (TAAAAA)12|D0|(AAAAAT)14
chr1 174902 174957 (A)1(AG)1(AAAG)1 55 +|+|+ (A)31|D0|(AG)13|D-2|(AGAA)15
chr1 180444 180472 (ACCCTG)1(AACCCT)1 28 +|+ (CCCTGA)16|D0|(TAACCC)12
chr1 180504 180537 (ACCCTC)1(AACCCT)1 33 +|+ (CCTCAC)20|D0|(CCCTAA)15
chr1 180552 180590 (AACCCT)2(AACCC)1 38 +|+|+ (AACCCT)14|D0|(AACCCT)14|D-2|(AACCC)12
chr1 180748 180912 (AACCCT)4 164 +|+|+|+ (CCCTAA)55|D0|(ACCCTA)44|D-2|(AACCCT)21|D-7|(AACCCT)44
Expects: INTEGER
Default: 0
Maximum distance between two individual repeats of a compound repeat. To report repeats overlapping by X bp use negative numbers i.e., -X. The default is 0bp.
Expects: FILE
Default: None
Input a genomic feature file to annotate the repeats in the genomic context.
Looper accepts both GFF and GTF format genomic feature files. Each repeat is
annotated w.r.t the closest gene and classified either as Genic, Exonic,
Intronic and Intergenic according to the position of the repeat. Besides this,
the repeat is also checked if it falls in the promoter region of the gene.
Annotation adds 7 columns to the default looper output which already consist 8
columns.
| S.No | Column | Description |
|---|---|---|
| 9 | Gene name | Name of the closest gene |
| 10 | Gene Start | Start position of gene in the Chromosome |
| 11 | Gene Stop | End position of gene in the Chromosome |
| 12 | Strand | The strand orientation of the gene |
| 13 | Genomic annotation | Annotation of the repeat w.r.t to the gene. Possible annotations are {Genic, Exonic, Intronic, Intergenic} |
| 14 | Promoter annotation | If repeat falls in the promoter region of the closest gene. The default promoter region is 1Kb upstream and downstream of TSS. |
| 15 | Distance from TSS | Distance of the repeat from the TSS of the gene. |
Expects: STRING
Default: GFF
Option to specify the format of the input genomic feature file. Accepted inputs
are GFF or GTF. More details about the GFF and GTF formats can be found
here.
Expects: STRING
Default: gene
The attribute key used for the name of the gene in the GFF/GTF file. In the
below example GFF file, we have the location of a gene and it's mRNA and exon
locations. The last column of the file specifies attributes associated with each
feature, like ID, Parent, gene etc. Looper uses on of the attribute to identify
the gene and also it's exons. In th below example the key "gene" can be used to
identify gene and the exons of the gene as they have the same gene name. Please
check your GFF/GTF file for a robust attribute key which can identify all genes
and their corresponding exons. We are actively working on better annotation
where we can identify genes and their exons based on the ID and Parent.
# Sample GFF
NC_004354.4 RefSeq gene 124370 126714 . - . ID=gene1;Name=CG17636;gbkey=Gene;gene=CG17636;gene_biotype=protein_coding;gene_synonym=DmelCG17636,EG:23E12.1;
NC_004354.4 RefSeq mRNA 124370 126714 . - . ID=rna1;Parent=gene1;Name=NM_001103384.3;gbkey=mRNA;gene=CG17636;transcript_id=NM_001103384.3
NC_004354.4 RefSeq exon 126626 126714 . - . ID=id13;Parent=rna1;gbkey=mRNA;gene=CG17636;transcript_id=NM_001103384.3
NC_004354.4 RefSeq exon 125495 126259 . - . ID=id14;Parent=rna1;gbkey=mRNA;gene=CG17636;transcript_id=NM_001103384.3
Expects: INT
Default: 1000
Upstream distance(bp) from the TSS of the gene to be considered as promoter
region. Default 1000.
Expects: INT
Default: 1000
Downstream distance(bp) from the TSS of the gene to be considered as promoter
region. Default 1000.
For queries or suggestions, please contact:
Divya Tej Sowpati - tej@ccmb.res.in
Akshay Kumar Avvaru - avvaru@ccmb.res.in