celescope 1.1.1

GEXSCOPE Single cell analysis

CeleScope

Requirements

• conda
• linux
• minimum 32GB RAM(to run STAR aligner)

Installation

git clone https://github.com/zhouyiqi91/CeleScope.git
cd CeleScope
conda env create -f celescope.yml

Reference genome

Homo sapiens

wget ftp://ftp.ensembl.org/pub/release-99/fasta/homo_sapiens/dna/Homo_sapiens.GRCh38.dna.primary_assembly.fa.gz
wget ftp://ftp.ensembl.org/pub/release-99/gtf/homo_sapiens/Homo_sapiens.GRCh38.99.gtf.gz

mkdir -p references/Homo_sapiens/Ensembl/GRCh38
gzip -c -d Homo_sapiens.GRCh38.dna.primary_assembly.fa.gz > references/Homo_sapiens/Ensembl/GRCh38/Homo_sapiens.GRCh38.fa
gzip -c -d Homo_sapiens.GRCh38.99.gtf.gz > references/Homo_sapiens/Ensembl/GRCh38/Homo_sapiens.GRCh38.99.gtf

conda activate celescope1.1

gtfToGenePred -genePredExt -geneNameAsName2 references/Homo_sapiens/Ensembl/GRCh38/Homo_sapiens.GRCh38.99.gtf /dev/stdout | \
    awk '{print $12"\t"$1"\t"$2"\t"$3"\t"$4"\t"$5"\t"$6"\t"$7"\t"$8"\t"$9"\t"$10}' > references/Homo_sapiens/Ensembl/GRCh38/Homo_sapiens.GRCh38.99.refFlat

STAR \
    --runMode genomeGenerate \
    --runThreadN 6 \
    --genomeDir references/Homo_sapiens/Ensembl/GRCh38 \
    --genomeFastaFiles references/Homo_sapiens/Ensembl/GRCh38/Homo_sapiens.GRCh38.fa \
    --sjdbGTFfile references/Homo_sapiens/Ensembl/GRCh38/Homo_sapiens.GRCh38.99.gtf \
    --sjdbOverhang 100

Mus musculus

wget ftp://ftp.ensembl.org/pub/release-99/fasta/mus_musculus/dna/Mus_musculus.GRCm38.dna.primary_assembly.fa.gz
wget ftp://ftp.ensembl.org/pub/release-99/gtf/mus_musculus/Mus_musculus.GRCm38.99.gtf.gz

mkdir -p references/Mus_musculus/Ensembl/GRCm38
gzip -c -d Mus_musculus.GRCm38.dna.primary_assembly.fa.gz > references/Mus_musculus/Ensembl/GRCm38/Mus_musculus.GRCm38.fa
gzip -c -d Mus_musculus.GRCm38.99.gtf.gz > references/Mus_musculus/Ensembl/GRCm38/Mus_musculus.GRCm38.99.gtf

conda activate celescope1.1

gtfToGenePred -genePredExt -geneNameAsName2 references/Mus_musculus/Ensembl/GRCm38/Mus_musculus.GRCm38.99.gtf /dev/stdout | \
    awk '{print $12"\t"$1"\t"$2"\t"$3"\t"$4"\t"$5"\t"$6"\t"$7"\t"$8"\t"$9"\t"$10}' > references/Mus_musculus/Ensembl/GRCm38/Mus_musculus.GRCm38.99.refFlat

STAR \
    --runMode genomeGenerate \
    --runThreadN 6 \
    --genomeDir references/Mus_musculus/Ensembl/GRCm38 \
    --genomeFastaFiles references/Mus_musculus/Ensembl/GRCm38/Mus_musculus.GRCm38.fa \
    --sjdbGTFfile references/Mus_musculus/Ensembl/GRCm38/Mus_musculus.GRCm38.99.gtf \
    --sjdbOverhang 100

Usage

Example

conda activate celescope1.1
python3 {CeleScope_path}/celescope.py rna\   
 --fq1 ./data/R2005212_L1_1.fq.gz\
 --fq2 ./data/R2005212_L1_2.fq.gz\
 --chemistry scopeV2.0.1\
 --genomeDir /SGR/references/Homo_sapiens/Ensembl/GRCh38\
 --sample R2005212\
 --thread 4

--fq1 gzipped FASTQ read 1 file path
--fq2 gzipped FASTQ read 2 file path
--chemistry chemistry version, choices=['scopeV2.0.0', 'scopeV2.0.1', 'scopeV2.1.0', 'scopeV2.1.1']
--genomeDir reference genome directory path
--sample sample name
--thread number of threads