epiAneufinder is an algorithm used for calling Copy Number Variations (CNVs) from single-cell ATAC (scATAC) data. Single-cell open chromatin profiling via the single-cell Assay for Transposase-Accessible Chromatin using sequencing (scATAC-seq) assay has become a mainstream measurement of open chromatin in single-cells. epiAneufinder exploits the read count information from scATAC-seq data to extract genome-wide copy number variations (CNVs) for each individual cell. epiAneufinder allows the addition of single-cell CNV information to scATAC-seq data, without the need of additional experiments, unlocking a layer of genomic variation which is otherwise unexplored.
Ramakrishnan, A., Symeonidi, A., Hanel, P. et al. epiAneufinder identifies copy number alterations from single-cell ATAC-seq data. Nat Commun 14, 5846 (2023). https://doi.org/10.1038/s41467-023-41076-1
All additional scripts used for the publication can be found here https://github.com/colomemaria/epiAneufinder_analyses.git
The algorithm works in three steps:
- Data preparation (binning, GC correction, removal of blacklisted regions)
- Genome segmentation based on maximum Anderson-Darling distance
- Gain/loss assignments
epiAneufinder is an R package
Installation example:
*OPTIONAL: Create a conda enviroment and install R and devtools
conda create -n epianeufinder r-base r-essentials
- Start R and install dependencies
install.packages(c("devtools", "BiocManager", "ggdendro"))
BiocManager::install(c("GenomicAlignments", "SummarizedExperiment", "plyranges", "Rsamtools", "GenomeInfoDb", "BSgenome.Hsapiens.UCSC.hg38", "GenomicRanges", "Biostrings", "BiocGenerics", "S4Vectors", "GenomicFeatures"))
- Load devtools and install the epiAneufinder package
library(devtools)
install_github("colomemaria/epiAneufinder")
The user should also download the blacklisted regions of the hg38 genome and update the parameter accordingly. By default, the genome version to be used is the hg38. If there is another genome/species that needs to be used, it should be installed by the user, along with the corresponding blacklisted regions.
epiAneufinder has been extensovely tested for the following version of dependencies: GenomicAlignments v1.28.0, SummarizedExperiment v1.22.0, plyranges v1.12.1, Rsamtools v2.8.0, GenomeInfoDb v1.28.4, BSgenome.Hsapiens.UCSC.hg38 v1.4.3, GenomicRanges v1.44.0, Biostrings v2.60.2, BiocGenerics v0.38.0, S4Vectors v0.30.0, GenomicFeatures v1.44.2, devtools v2.4.3, BiocManager v1.30.16 and ggdendro v0.1.22.
The installation process of epiAneufinder, in a prepared enviroment (all dependencies already installed), takes approximatelly 3 minutes.
The software has been tested for Linux and MacOS.
This is a short introduction in the epiAneufinder functionality, more information can be found in the vignette introduction-epiAneufinder.
library(epiAneufinder)
epiAneufinder(input="sample.tsv", #Enter path to your fragments.tsv file or the folder containing bam files
outdir="epiAneufinder_results", #Path to the directory where results should be written
blacklist="hg38-blacklist.v2.bed", #Path to bed file that contains the blacklisted regions of your genome
windowSize=1e5,
genome="BSgenome.Hsapiens.UCSC.hg38", #Substitute with relevant BSgenome
exclude=c('chrX','chrY','chrM'),
reuse.existing=TRUE,
title_karyo="Karyogram of sample data",
ncores=4,
minFrags=20000,
minsizeCNV=0,
k=4,
plotKaryo=TRUE)
We also provide a wrapper script to be used (epiAneufinder_wrapper.R).
The user needs to update the path/name of the input data folder, as well as the path/name of the blacklisted regions to use on the wrapper. Test data and the hg38 blacklisted regions can be found in the sample_data folder.
The user may also change on the wrapper any of the parameters of the algorithm, if needed.
Once the user has updated the parameters on the wrapper, the wrapper can be called from console with the following command:
Rscript epiAneufinder_wrapper.R
Depending on the available resources, running the demo shouldn't take more than 15 minutes. Tested in a typical desktop, running time was 7.34 minutes with 6Gb of RAM used.
Contributors names and contact info
Akshaya Ramakrishnan (akshaya4r@gmail.com)
Aikaterini Symeonidi (aikaterini.symeonidi@helmholtz-muenchen.de)
Patrick Hanel (patrick.hanel@helmholtz-muenchen.de)
Katharina Schmid (katharina.schmid@bmc.med.lmu.de)
Maria Richter (maria.richter@bmc.med.lmu.de)
Michael Schubert
Maria Colomé-Tatché (maria.colome@helmholtz-muenchen.de)
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1.0.3
- Added the option to start the algorithm directly from a count matrix (Warning: performance might drop compared to bam/fragment files, see vignette)
- Extended the text output to show a few more infos, e.g. number cells and windows
- Added the parameter mapqFilter for a flexible filter of the bam files
- Added a function to split cells into sublcones
- Added to additional plotting functions, to plot the read distribution of an individual cell and to plot additional annotations in a side bar next to the karyogram
- Added a vignette to improve the documentation
-
1.0.2
- Updated function documentations
- Included a better explaining error message for mismatching chromosome versions
- Corrected minor bug if one chromosome fits in one window
- Made karyogram plotting optional (argument plotKaryo)
- Added in the wrapper the default parameters as used in the paper
-
0.1
- Initial Release
This project is licensed under the GNU General Public License v3.0 - see the LICENSE.md file for details