This repository is home of R package uncoverappLib launching a *unCOVERApp*,
a web application for clinical assessment and annotation of coverage gaps in
target genes. Read more about unCOVERApp on [biorxiv](https://www.biorxiv.org/content/10.1101/2020.02.10.939769v1)
[](https://zenodo.org/badge/latestdoi/254597958)
# Table of content
* [Prerequisites](#Prerequisites)
* [Installation](#Installation)
* [Introduction](#Introduction)
* [Download_annotation_files](#Download_annotation_files)
* [Input](#Input)
* [Usage](#Usage)
## Prerequisites
This app requires following dependencies:
- R >= 4.0.0
- java installed
- annotation files (`sorted.bed.gz` and `sorted.bed.gz.tbi`) that can be
downloaded on [Zenodo](https://zenodo.org/record/3747448#.XpBmnVMzbOR) with
`getAnnotationFiles()` function of *uncoverappLib*.
## Installation
To install this package, start R and enter:
``` {r}
if (!requireNamespace("BiocManager", quietly = TRUE))
install.packages("BiocManager")
#BiocManager::install("uncoverappLib")
```
The development version can be installed directly from Github:
``` {r}
install.packages("devtools")
devtools::install_github("Manuelaio/uncoverappLib")
```
Major informations about unCOVERApp R dependences are
[here](https://github.com/Manuelaio/test_dependence) .
[](https://travis-ci.com/Manuelaio/test_dependence)
## Introduction
The rapid spread of NGS-technology has been coupled since its beginning with
development of bioinformatic tools for data analysis and interpretation.
However, despite increasing accuracy of available approaches, the need to
assess sequencing quality of the analysis targets at the base-pair resolution
poses growing challenges especially in the clinical settings.
In diagnostics indeed, meticulous investigation of every single target base is
often required to exclude that pathogenic events across the gene of interest
may be missed due to uneven sequence coverage.
**unCOVERApp** is an interactive web-application
for graphical inspection of sequence coverage within gene regions.
unCOVERApp highlights low coverage genomic positions, according to the coverage
threshold specified by the user, providing *dbNSFP-based annotation*s for
clinical assessment of low coverage regions.
It implements basic statistical tools such as binomial probability calculation
that genomic positions are adequately
covered, and
[maximum credible allele frequency](http://cardiodb.org/allelefrequencyapp/).
# Download_annotation_files
To associate low-coverage sites with functional and clinical annotations,
unCOVERApp uses [dbNSFP](https://sites.google.com/site/jpopgen/dbNSFP)
version 4.0 stored in two file:
* `sorted.bed.gz`: a genomically-sorted, TABIX-indexed, BGZipped BED file
containing selected columns from dbNSFP version v4.0.
* `sorted.bed.gz.tbi`: TABIX-indexed file.
Those files are stored on Zenodo at following
[link](https://zenodo.org/record/3747448#.XpBmnVMzbOR) for downloading.
*sorted.bed.gz* encloses prediction scores (MutationAssessor, SIFT, CADD,
M-CAP, Polyphen2-HVAR), allele frequencies observed in
gnomAD data, dbsnp accession number, HGVS notations and clinical annotation
information from ClinVar and OMIM. Loading *sorted.bed.gz* allows the annotation
of each low coverage genomic position user-defined. .
Run following commands to correctly create a unique cache for **uncoverappLib**
using **BiocFileCache** package.
``` {r}
library(uncoverappLib)
getAnnotationFiles()
```
# Input
As input file unCOVERApp takes:
- a text file, with .txt extension, containing HGNC official gene name one per
row
- a text file, with .list extension, containing absolute paths to BAM files
(one per row). In the output file, the first written bam file correspond to
sample 1 and so forth.
For more details on working with unCOVERApp see Vignette.
# Usage
Load library and set up R environment with annotation file as following.
The way to launch unCOVERApp is *run.uncoverapp(where="window")* function.
``` {r}
library(uncoverappLib)
run.uncoverapp(where="window")
```
User can define where uncoverapp will be launched wiht `where` option:
- `browser` option will open `uncoverapp` in your default browser
- `viewer` option will open `uncoverapp` in RStudio viewer
- `window` option will open `uncoverapp` in RStudio RStudio
For more details on working with uncoverapp see Vignette or [Documentation.pdf](https://github.com/Manuelaio/unCOVERApp/blob/master/Documentation.pdf) on Github.
