KITSUNE is a toolkit for evaluating the length of k-mer in a given genome dataset for alignment-free phylogenomic analysis.
K-mer based approach is simple and fast yet has been widely used in many applications including biological sequence comparison. However, selection of an appropriate k-mer length to obtain good information content for comparison is normally overlooked. The optimum k-mer length is a prerequisite to obtain biological meaningful genomic distance for assessment of phylogenetic relationships. Therefore, we have developed KITSUNE to aid k-mer length selection process in a systematic way, based on a three-steps approach described in Viral Phylogenomics Using an Alignment-Free Method: A Three-Step Approach to Determine Optimal Length of k-mer.
KITSUNE will calculate the three matrices across considered k-mer range:
- Cumulative Relative Entropy (CRE)
- Average number of Common Features (ACF)
- Observed Common Features (OCF)
Moreover, KITSUNE also provides various genomic distance calculations from the k-mer frequency vectors that can be used for species identification or phylogenomic tree construction.
Note: If you use KITSUNE in your research, please cite: KITSUNE: A Tool for Identifying Empirically Optimal K-mer Length for Alignment-free Phylogenomic Analysis
Kitsune is developed under python version 3 environment. We recommend users use python >= v3.5.
Requirement packages: scipy >= 0.18.1, numpy >= 1.1.0, tqdm >= 4.32
Kitsune also requires Jellyfish for k-mer counting as an external software dependency. Thus, you need to install it before running the tool: https://github.com/gmarcais/Jellyfish
pip install kitsune# Clone the GitHub repository
git clone https://github.com/natapol/kitsune
# Move to the kitsune folder
cd kitsune/
# Install
python setup.py installcommand for listing help
$ kitsune --help
usage: kitsune <command> [<args>]
Available commands:
acf Compute average number of common features between signatures
cre Compute cumulative relative entropy
dmatrix Compute distance matrix
kopt Compute recommended choice (optimal) of kmer within a given kmer interval for a set of genomes using the cre, acf and ofc
ofc Compute observed feature frequencies
Use --help in conjunction with one of the commands above for a list of available options (e.g. kitsune acf --help)Kitsune provides three commands to calculate an appropiate k-mer using CRE, ACF, and OCF:
$ kitsune cre -h
usage: kitsune (cre) [-h] --filename FILENAME [--fast] [--canonical] -ke KEND
[-kf KFROM] [-t THREAD] [-o OUTPUT]
Calculate k-mer from cumulative relative entropy of all genomes
optional arguments:
-h, --help show this help message and exit
--filename FILENAME A genome file in fasta format (default: None)
--fast Jellyfish one-pass calculation (faster) (default:
False)
--canonical Jellyfish count only canonical mer (default: False)
-ke KEND, --kend KEND
Last k-mer (default: None)
-kf KFROM, --kfrom KFROM
Calculate from k-mer (default: 4)
-t THREAD, --thread THREAD
-o OUTPUT, --output OUTPUT
Output filename (default: None)$ kitsune acf -h
usage: kitsune (acf) [-h] --filenames FILENAMES [FILENAMES ...] [--fast]
[--canonical] -k KMERS [KMERS ...] [-t THREAD]
[-o OUTPUT]
Calculate an average number of common features pairwise between one genome
against others
optional arguments:
-h, --help show this help message and exit
--filenames FILENAMES [FILENAMES ...]
Genome files in fasta format (default: None)
--fast Jellyfish one-pass calculation (faster) (default:
False)
--canonical Jellyfish count only canonical mer (default: False)
-k KMERS [KMERS ...], --kmers KMERS [KMERS ...]
Have to state before (default: None)
-t THREAD, --thread THREAD
-o OUTPUT, --output OUTPUT
Output filename (default: None)$ kitsune ofc -h
usage: kitsune (ofc) [-h] --filenames FILENAMES [FILENAMES ...] [--fast]
[--canonical] -k KMERS [KMERS ...] [-t THREAD]
[-o OUTPUT]
Calculate an observe feature frequency
optional arguments:
-h, --help show this help message and exit
--filenames FILENAMES [FILENAMES ...]
Genome files in fasta format (default: None)
--fast Jellyfish one-pass calculation (faster) (default:
False)
--canonical Jellyfish count only canonical mer (default: False)
-k KMERS [KMERS ...], --kmers KMERS [KMERS ...]
-t THREAD, --thread THREAD
-o OUTPUT, --output OUTPUT
Output filename (default: None)kitsune cre --filename genome1.fna -kf 5 -ke 10
kitsune acf --filenames genome1.fna genome2.fna -k 5
kitsune ofc --filenames genome_fasta/* -k 5Kitsune provides a commands to calculate genomic distance using different distance estimation method. Users can assess the impact of a selected k-mer length on the genomic distnace of choice below.
| distance option | name |
|---|---|
| braycurtis | Bray-Curtis distance |
| canberra | Canberra distance |
| chebyshev | Chebyshev distance |
| cityblock | City Block (Manhattan) distance |
| correlation | Correlation distance |
| cosine | Cosine distance |
| euclidean | Euclidean distance |
| jensenshannon | Jensen-Shannon distance |
| sqeuclidean | Squared Euclidean distance |
| dice | Dice dissimilarity |
| hamming | Hamming distance |
| jaccard | Jaccard-Needham dissimilarity |
| kulsinski | Kulsinski dissimilarity |
| rogerstanimoto | Rogers-Tanimoto dissimilarity |
| russellrao | Russell-Rao dissimilarity |
| sokalmichener | Sokal-Michener dissimilarity |
| sokalsneath | Sokal-Sneath dissimilarity |
| yule | Yule dissimilarity |
| mash | MASH distance |
| jsmash | MASH Jensen-Shannon distance |
| jaccarddistp | Jaccard-Needham dissimilarity Probability |
| euclidean_of_frequency | Euclidean distance of Frequency |
Kitsune provides a choice of distance transformation proposed by Fan et.al.
$ kitsune dmatrix -h
usage: kitsune (dmatrix) [-h] [--filenames [FILENAMES [FILENAMES ...]]]
[--fast] [--canonical] -k KMER [-i INPUT] [-o OUTPUT]
[-t THREAD] [--transformed]
[-d {braycurtis,canberra,jsmash,chebyshev,cityblock,correlation,cosine,dice,euclidean,hamming,jaccard,kulsinsk,matching,rogerstanimoto,russellrao,sokalmichener,sokalsneath,sqeuclidean,yule,mash,jaccarddistp}]
[-f FORMAT]
Calculate a distance matrix
optional arguments:
-h, --help show this help message and exit
--filenames [FILENAMES [FILENAMES ...]]
Genome files in fasta format (default: None)
--fast Jellyfish one-pass calculation (faster) (default:
False)
--canonical Jellyfish count only canonical mer (default: False)
-k KMER, --kmer KMER
-i INPUT, --input INPUT
List of genome files in txt (default: None)
-o OUTPUT, --output OUTPUT
Output filename (default: None)
-t THREAD, --thread THREAD
--transformed
-d {braycurtis,canberra,jsmash,chebyshev,cityblock,correlation,cosine,dice,euclidean,hamming,jaccard,kulsinsk,matching,rogerstanimoto,russellrao,sokalmichener,sokalsneath,sqeuclidean,yule,mash,jaccarddistp}, --distance {braycurtis,canberra,jsmash,chebyshev,cityblock,correlation,cosine,dice,euclidean,hamming,jaccard,kulsinsk,matching,rogerstanimoto,russellrao,sokalmichener,sokalsneath,sqeuclidean,yule,mash,jaccarddistp}
-f FORMAT, --format FORMATExample of choosing distance option:
kitsune dmatrix --filenames genome1.fna genome2.fna -k 11 -d jaccard --canonical --fast -o output.txt
kitsune dmatrix --filenames genome1.fna genome2.fna -k 11 -d hensenshannon --canonical --fast -o output.txtKitsune provides a wrap-up comand to find optimum k-mer length for a given set of genome within a given kmer interval.
$ kitsune kopt -h
usage: kitsune (kopt) [-h] [--acf-cutoff ACF_CUTOFF] [--canonical]
[--closely-related] [--cre-cutoff CRE_CUTOFF] [--fast]
--filenames FILENAMES [--hashsize HASHSIZE]
[--in-memory] [--k-min K_MIN] --k-max K_MAX
[--lower LOWER] [--nproc NPROC] [--output OUTPUT]
[--threads THREADS]
Optimal kmer size selection for a set of genomes using Average number of
Common Features (ACF), Cumulative Relative Entropy (CRE), and Observed Common
Features (OCF). Example: kitsune kopt --filenames genomeList.txt --k-min 4
--k-max 12 --canonical --fast
optional arguments:
-h, --help show this help message and exit
--acf-cutoff ACF_CUTOFF
Cutoff to use in selecting kmers whose ACFs are >=
(cutoff * max(ACF)) (default: 0.1)
--canonical Jellyfish count only canonical kmers (default: False)
--closely-related Use in case of closely related genomes (default:
False)
--cre-cutoff CRE_CUTOFF
Cutoff to use in selecting kmers whose CREs are <=
(cutoff * max(CRE)) (default: 0.1)
--fast Jellyfish one-pass calculation (faster) (default:
False)
--filenames FILENAMES
Path to the file with the list of genome files paths.
There should be at list 2 input genomes (default:
None)
--hashsize HASHSIZE Jellyfish initial hash size (default: 100M)
--in-memory Keep Jellyfish counts in memory (default: False)
--k-min K_MIN Minimum kmer size (default: 4)
--k-max K_MAX Maximum kmer size (default: None)
--lower LOWER Do not let Jellyfish output kmers with count < --lower
(default: 1)
--nproc NPROC Maximum number of CPUs to make it parallel (default:
1)
--output OUTPUT Path to the output file (default: None)
--threads THREADS Maximum number of threads for Jellyfish (default: 1)First download the example files. Download
kitsune kopt --filenames genome_list --k-min 6 --k-max 21 --canonical --fast --threads 4 --nproc 2 --output out.txt
⚠️ Please be aware that this command will use big computational resources when large number of genomes and/or large genome size are used as the input.