FiTnEss

Finding Tn-Seq Essential genes (FiTnEss)

Poster: FiTnEss_poster.pdf

FiTnEss is an R package using Transposon insertion sequencing data to identify essential genes in the genome.

Original paper on bioRxiv: Defining the core essential genome of Pseudomonas aeruginosa

Publication: Defining the core essential genome of Pseudomonas aeruginosa

Quick start

After installing FiTnEss package, run main FiTnEss function by FiTnEss_Run

Function arguments

Arguments in this function include:

• strain
• file_location: path and name of tally file for run: e.g. "/home/your_folder/your_tally.txt"
• permissive_file: path and name of non-permissive TA site file that generated from genomic pre-processing step: e.g. "/home/your_folder/non_permissive_TA_sites.txt"
• homologous_file: path and name of homologous TA site file that generated from pre-processing step: e.g. "/home/your_folder/homologous_TA_sites.txt"
• gene_file: path and name of GFF3 gene annotation file. For example, GFF3 file could be downloaded from Pseudomonas Genome Database: e.g. "/your/folder/location/your_gff3_file.txt"
• save_location: path and name of where to save final results file: e.g. "/home/results_folder/results.xlsx"
• repeat_time: how many times to run the pipeline in order to obtain best results: by default, we run 3 times.

Step 1. install devtools package

install.packages("devtools")

Step 2. install FiTnEss package from github

devtools::install_github("ruy204/FiTnEss")

Step 3. load FiTnEss and dependent packages

Packages <- c("dplyr","fBasics","goftest","openxlsx","scales","stats","tidyr")
lapply(Packages, library, character.only = TRUE)

require(FiTnEss)

Step 4. run FiTnEss

FiTnEss_Run("PA14",
            "/your/folder/location/Test_set_P_aeruginosa/sample_data/PA14_M9_rep1_tally.txt",
            "/your/folder/location/Test_set_P_aeruginosa/TAsite_info/nonpermissive_TA_sites.txt",
            "/your/folder/location/Test_set_P_aeruginosa/TAsite_info/homologous_TA_sites.txt",
            "/your/folder/location/Test_set_P_aeruginosa/genome_info/PA14_gff.txt",
            "/your/folder/location/Test_set_P_aeruginosa/sample_data/test_results.xlsx",
            repeat_time = 3)

Step 5. retrieve results

Locus.CIA	gtot	Nta	pvalue	padj	Ess_fwer	pfdr	Ess_fdr
PA14_00410	5	1	0.015989	1	NE_fwer	0.093033	NE_fdr

Each tab in the .xlsx file saves results from each replicate. Within each results table, there are 8 columns:

• Locus.CIA: gene index
• gtot: total reads for the gene
• Nta: number of TA sites in this gene
• pvalue: unadjusted p-value of being essential
• padj: FWER-adjusted p-value
• Ess_fwer: confident essential category
• pfdr: FDR-adjusted p-value
• Ess_fdr: candidate essential category