Decoil (deconvolve extrachromosomal circular DNA isoforms from long-read data) is a software package for reconstruction circular DNA.
As a prequisite you need to have install docker or singularity (you can install this from the official website or using conda).
Download decoil docker image from docker-hub. This contains all the dependencies needed to run the software. No additional installation needed. All the environment, packages, dependencies are all specified in the docker/singularity image.
# docker
docker pull madagiurgiu25/decoil:1.1.2-slim# singularity
singularity pull decoil.sif docker://madagiurgiu25/decoil:1.1.2-slimTo test your installation check the Example.
To run Decoil on your data you need to cofigure the following parameters:
# run decoil with your input with standard parameters
BAM_INPUT="<absolute path to your BAM file>"
OUTPUT_FOLDER="<absolute path to your output folder>"
NAME="<sample name>"
GENOME="<absolute path to your reference genome file>"
ANNO="<absolute path to your gtf annotation file>"and then run the following command:
# docker
docker run -it --platform=linux/amd64 \
-v ${BAM_INPUT}:/data/input.bam \
-v ${BAM_INPUT}.bai:/data/input.bam.bai \
-v ${GENOME}:/annotation/reference.fa \
-v ${ANNO}:/annotation/anno.gtf \
-v ${OUTPUT_FOLDER}:/mnt \
-t madagiurgiu25/decoil:1.1.2-slim \
decoil-pipeline sv-reconstruct \
-b /data/input.bam \
-r /annotation/reference.fa \
-g /annotation/anno.gtf \
-o /mnt --name ${NAME}# singularity
mkdir -p ${OUTPUT_FOLDER}
mkdir -p ${OUTPUT_FOLDER}/logs
mkdir -p ${OUTPUT_FOLDER}/tmp
singularity run \
--bind ${OUTPUT_FOLDER}/logs:/mnt/logs \
--bind ${OUTPUT_FOLDER}/tmp:/tmp \
--bind ${BAM_INPUT}:/data/input.bam \
--bind ${BAM_INPUT}.bai:/data/input.bam.bai \
--bind ${GENOME}:/annotation/reference.fa \
--bind ${ANNO}:/annotation/anno.gtf \
--bind ${OUTPUT_FOLDER}:/mnt \
decoil.sif \
decoil-pipeline sv-reconstruct \
-b /data/input.bam \
-r /annotation/reference.fa \
-g /annotation/anno.gtf \
-o /mnt --name ${NAME}You can install the latest version of Decoil repository (git and conda/mamba required):
git clone https://github.com/madagiurgiu25/decoil-pre.git
cd decoil-pre
# create conda environment
mamba env create -f environment.yml
conda activate envdecoil
python -m pip install -r requirements.txt
python setup.py build install
# add decoil in $PATH
ROOT=`dirname $(which decoil)`
export PATH=$PATH:$ROOTAnd check if the installation worked:
# might take a while
decoil-pipeline --version
decoil --versionAn overview about the available functionalities:
| decoil-pipeline | decoil | decoil-viz | |
|---|---|---|---|
| (recommended) | (advanced users) | (recommended) | |
| SV calling | x | ||
| coverage track | x | ||
| reconstruction | x | x | |
| visualization | x | ||
| docker | x | x | x |
| singularity | x | x | x |
decoil-pipeline (recommended)To reconstruct ecDNA we recommend to use decoil-pipeline using the sv-reconstruct mode.
This requires only a .bam file as input and generates internally all the files required for the reconstruction.
# call help
docker run -it --platform=linux/amd64 -t madagiurgiu25/decoil:1.1.2-slim decoil-pipeline --help
usage: decoil-pipeline <workflow> <parameters> [<target>]
Example:
# run decoil including the processing and visualization steps
decoil-pipeline -f sv-recontruct --bam <input> --outputdir <outputdir> --name <sample> --sv-caller <sniffles> -r <reference-genome> -g <annotation-gtf>
Decoil 1.1.2: reconstruct ecDNA from long-read data
positional arguments:
{sv-only,sv-reconstruct,reconstruct-only}
sub-command help
sv-only Perform preprocessing
sv-reconstruct Perform preprocessing and reconstruction
optional arguments:
-h, --help show this help message and exit
--version show program's version number and exit
-n, --dry-run
-f, --force
-c, --use-condaThe pipeline has the following running modes:
sv-onlysv-reconstructreconstruct-onlydecoil-viz (recommended)To interpret and visualize the results of the ecDNA reconstruction threads, use decoil-viz.
decoil (advanced users only)This configuration is the most flexible and allows users to use their own SV calls. For details go here.
Check recommendations for filtering or debugging in the FAQ section.
The relevant output files for the users are:
reconstruct.bed - contains all genomic fragments composing all reconstructionsreconstruct.ecDNA.bed - contains all genomic fragments composing all the ecDNA labeled reconstructionssummary.txt - summarize all the circular reconstructionscat reconstruct.bed
#chr start end circ_id fragment_id strand coverage estimated_proportions
chr2 15585356 15633376 0 5 + 149 75
chr3 11150000 11160001 0 41 - 103 75
chr3 11049997 11060001 0 33 + 117 75
chr2 15585356 15633376 3 5 + 149 36
chr3 11150000 11160001 3 41 - 103 36
chr3 11049997 11060001 3 33 + 117 36
chr2 15585356 15633376 3 5 + 149 36
chr2 16521052 16628305 3 13 + 37 36
chr3 10981202 11028470 3 25 - 31 36
chr12 68807722 68970910 2 53 + 252 252
cat summary.txt
circ_id chr_origin size(MB) label topology_idx topology_name estimated_proportions
0 chr3,chr2 0.068025 4 multi_region_inter_chr 75
3 chr3,chr2 0.270566 ecDNA 5 simple_duplications 36
2 chr12 0.163188 ecDNA 0 simple_circle 252If you use Decoil for your work please cite our pre-print:
Madalina Giurgiu, Nadine Wittstruck, Elias Rodriguez-Fos, Rocio Chamorro Gonzalez, Lotte Bruckner, Annabell Krienelke-Szymansky, Konstantin Helmsauer, Anne Hartebrodt, Richard P. Koche, Kerstin Haase, Knut Reinert, Anton G. Henssen. Decoil: Reconstructing extrachromosomal DNA structural heterogeneity from long-read sequencing data. bioRxiv, 2023, DOI: https://doi.org/10.1101/2023.11.15.567169
Decoil is distributed under the BSD 3-Clause license. Consult the accompanying LICENSE file for more details.
Decoil and the content of this research-repository (i) is not suitable for a medical device; and (ii) is not intended for clinical use of any kind, including but not limited to diagnosis or prognosis.