Variant Discovery    ◾    149

variants. The low-impact variants are the synonymous ones that do not change protein

behavior; however, they may still have some effects. Variants on introns (intron-variants)

and variants in the downstream region of a gene are annotated as MODIFIER since they

affect non-coding regions, where prediction of the impact is difficult or there is no substan-

tial evidence of any effect.

To use snpEff, we should install snpEff software and download the database of interest.

Installation of SnpEff software requires Java V1.8 or later installed on your computer. The

installation instructions are available at “https://pcingola.github.io/SnpEff/download/” or

“http://pcingola.github.io/SnpEff/”. First, download the compressed folder using “wget”

Linux command and then decompress it with “unzip” command. You can also install the

binary.

wget https://snpeff.blob.core.windows.net/versions/snpEff_latest_

core.zip

unzip snpEff_latest_core.zip

This will create the directory “snpEff”, which contains two Java executable files with “.jar”

extension (snpEff.jar and snpSift.jar), a snpEff configuration file (snpEff.config), a license

file, and four subdirectories (“examples”, “exec”, “galaxy”, and “script”). By default, the

database will be stored in a subdirectory “data” in the “snpEff” directory but we can change

it by opening “snpEff.config” and editing “data.dir = ./data/” to the path that we need.

The database can be downloaded automatically and this is recommended but you can

also install it manually. For example, if you need to download the human database manu-

ally, you can run the following:

java -jar snpEff.jar download GRCh38.

Figure 4.11 shows the directory structure of the snpEff. If you installed the binary files, you

can use any of the executables without the “java” command as follows:

snpEff download GRCh38.

When you use snpEff, you must provide the right file path. For instance, if you are just one

step out of the “snpEff” directory, then you can run:

FIGURE 4.11  snpEff directory structure.