# Introduction to TnT

## Motivation

A common task in bioinformatics is to create visualization of genomic data along genomic coordinates, together with necessary genomic annotation features like genes and transcripts on the same coordinate, in order to make sense of those data.

Typically, this can be accomplished with a browser-based genome browser like UCSC genome browser or IGV, which requires to export the data from R. There are also R packages developed to address this issue but using static graphs, e.g. Gviz and ggbio.

While bioconductor have packages that excel at representing and analyzing such genomic data, there lacks a flexible and interactive way to view them. Sometimes there is no need for a full-functional genome browser but a fast and convenient way to view the data which are typically represented by a R object. It should also be interactive to aid exploration, for example it may be dragable and it may enable tooltips to get detailed information about a separate feature quickly.

This is just the motivation of TnT: it aims to provide an interactive and flexible approach to visualize genomic data right in R. In order to accomplish this goal, TnT wraps the TnT javascript libraries and provides bindings to common bioconductor classes (e.g. GRanges, TxDb) that represent genomic data. The TnT javascript libraries which the R package is based on are a set of javascript libraries for visualizing trees- and track-based annotations, which can be used to create a simple genome browser.

TnT is a new package, any feedback or suggestion would appreciated, please email to Jialin Ma < marlin-@gmx.cn >. You can also find the source repository at https://github.com/marlin-na/TnT and the documentation site at http://tnt.marlin.pub . This vignette will also be extended in the future to include more details.

## Install

You can install the stable version of TnT from Bioconductor:

source("https://bioconductor.org/biocLite.R")
biocLite("TnT")

Or alternatively, install the devel version from github:

devtools::install_github("marlin-na/TnT")

Then attach the package.

suppressPackageStartupMessages(library(TnT))

This vignette will assume readers have experience with common data structures in bioconductor, especially GRanges class from GenomicRanges package.

## Track Constructors

Overall, the package works by constructing tracks from data (GRanges, TxDb, EnsDb, etc.), and then constructing a tnt board from a list of tracks.

So the first step is to choose a track constructor and use it to construct tracks from data. Different constructors have been provided by the package for different features and data types.

As a simple example, to construct a block track from GRanges object

gr <- GenomicRanges::GRanges("chr7",
ranges = IRanges(
start = c(26549019L, 26564119L, 26585667L, 26591772L, 26594192L, 26623835L,
26659284L, 26721294L, 26821518L, 26991322L),
end =   c(26550183L, 26564500L, 26586158L, 26593309L, 26594570L, 26624150L,
26660352L, 26721717L, 26823297L, 26991841L)),
ID = 1:10,
Name = paste("My Range", 1:10)
)
btrack <- TnT::BlockTrack(gr)
btrack
## A BlockTrack
## | Label: gr
## | Background:    missing, use 'white'
## | Height:    30
## | Data:  RangeTrackData object with 10 ranges and 3 metadata columns:
## |         seqnames               ranges strand |        tooltip       color
## |            <Rle>            <IRanges>  <Rle> |   <data.frame> <character>
## |     [1]     chr7 [26549019, 26550183]      * |   1:My Range 1        blue
## |     [2]     chr7 [26564119, 26564500]      * |   2:My Range 2        blue
## |     [3]     chr7 [26585667, 26586158]      * |   3:My Range 3        blue
## |     [4]     chr7 [26591772, 26593309]      * |   4:My Range 4        blue
## |     [5]     chr7 [26594192, 26594570]      * |   5:My Range 5        blue
## |     [6]     chr7 [26623835, 26624150]      * |   6:My Range 6        blue
## |     [7]     chr7 [26659284, 26660352]      * |   7:My Range 7        blue
## |     [8]     chr7 [26721294, 26721717]      * |   8:My Range 8        blue
## |     [9]     chr7 [26821518, 26823297]      * |   9:My Range 9        blue
## |    [10]     chr7 [26991322, 26991841]      * | 10:My Range 10        blue
## |               key
## |         <integer>
## |     [1]         1
## |     [2]         2
## |     [3]         3
## |     [4]         4
## |     [5]         5
## |     [6]         6
## |     [7]         7
## |     [8]         8
## |     [9]         9
## |    [10]        10
## |    -------
## |    seqinfo: 1 sequence from an unspecified genome; no seqlengths

As you can see, meta-columns of GRanges have been converted to the tooltip column in track data. This is the default argument behavior, see

args(TnT::BlockTrack)
## function (range, label = deparse(substitute(range)), tooltip = mcols(range),
##     color = "blue", background = NULL, height = 30)
## NULL

The tooltip can be given as a data frame parallel to the data, the color argument can also be a character vector parallel to the data setting colors for each individual range.

In order to view track, simply put that track into a TnTBoard/TnTGenome:

TnT::TnTGenome(btrack)
## - Missing argument view.range:
##   automatically select 26493666..27047193 on seqlevel chr7...
## - Missing argument coord.range and seqlength is unknown:
##   automatically set coordinate limit to 26454128..27086731 ...

You can drag to move, scroll to zoom and click on feature to see the tooltip.

Similarly, tracks of different features could be constructed with other constructors. Here is a table showing these constructors and their data sources. Links to examples of each track type are also provided and you are recommended to go through them.

Constructor Source Feature type Example
BlockTrack GRanges block Block Track
VlineTrack Width-one GRanges vline Vline Track
PinTrack Width-one GRanges paired with values pin Pin Track
LineTrack Width-one GRanges paired with values line Line and Area Track
AreaTrack Width-one GRanges paired with values area Line and Area Track
GeneTrackFromTxDb TxDb gene Gene Track and Feature Track
FeatureTrack GRanges gene Gene Track and Feature Track
GroupFeatureTrack GRangesList tx Tx Track and GroupFeatureTrack
TxTrackFromTxDb TxDb tx Tx Track and GroupFeatureTrack
TxTrackFromGRanges GRanges paired with ‘type’ and ‘tx_id’ tx Tx Track and GroupFeatureTrack
merge Two or more tracks composite Composite Track

It is worthwhile to mention CompositeTrack here: you can merge multiple tracks to construct a CompositeTrack so that different types of features can be shown within one track. See example here.

## Track Manipulation

Given a constructed track, we may want to access or modify its data and options.

There are three common options for all types of tracks, they are background, height and label. These three options can be accessed and modified via trackSpec and trackSpec<-. For example:

TnT::trackSpec(btrack, "background")
## NULL
btrack2 <- btrack
TnT::trackSpec(btrack2, "background") <- "blanchedalmond"
TnT::trackSpec(btrack2, "label")      <- "My Ranges"
TnT::trackSpec(btrack2, "height")     <- 50

Data of tracks are normally stored with a class that inherits GRanges (except CompositeTrack, in which the data is stored as a list of tracks), and can be accessed or modified via trackData or trackData<-. There are also convenience shortcuts track$name and track$name <- value for trackData(track)$name and trackData(track)$name <- value, respectively. As an example:

btrack2$color # Equivalent to trackData(btrack2)$color
##  [1] "blue" "blue" "blue" "blue" "blue" "blue" "blue" "blue" "blue" "blue"
btrack2$color <- "darkseagreen4" # Equivalent to trackData(btrack2)$color <- "darkseagreen4"

As an example, let’s also modify the data:

TnT::trackData(btrack2) <- GenomicRanges::shift(TnT::trackData(btrack2), 10000)

Finally, we put the modified track and the original track together to see the difference.

TnT::TnTBoard(list(btrack, btrack2))
## - Missing argument view.range:
##   automatically select 26504916..27045943 on seqlevel chr7...
## - Missing argument coord.range and seqlength is unknown:
##   automatically set coordinate limit to 26451985..27098874 ...

Another thing we may want to modify is tooltip. By constructing the track via constructors (except those constructed from TxDb), tooltip can be given as a data frame parallel to the data. After the track is constructed, the tooltip can accessed via tooltip(track) which is an shortcut to trackData(track)\$tooltip. For example:

TnT::tooltip(btrack2) <- cbind(TnT::tooltip(btrack2),
as.data.frame(TnT::trackData(btrack2)))
TnT::TnTGenome(btrack2, view.range = TnT::trackData(btrack2)[4] * .05)
## - Missing argument coord.range and seqlength is unknown:
##   automatically set coordinate limit to 26464128..27096731 ...

Try to click on the block to see the tooltip.

## TnTBoard and TnTGenome

In previous examples, we have already seen how to show tracks with a TnTBoard or TnTGenome. A TnTBoard stores a list of tracks and show them with the same coordinate. You may already have noticed the difference between TnTBoard and TnTGenome: TnTGenome is just a TnTBoard with axis and location label.

In this part, I will introduce some arguments that can be optionally provided to control the board. They are:

• view.range: GRanges, to set the initial view range.
• coord.range: IRanges or numeric, to set the cooordinate limit.
• zoom.allow: IRanges or numeric, to set the limit of extent when zooming in and out.
• allow.drag: Logical, if FALSE, the board will not be able to move or zoom.

In case that view.range, coord.range and zoom.allow not provided, TnT will take a guess on them. Some considerations are:

• view.range: Try to use the seqlevel on which all tracks have features and try to use intersection of ranges of all tracks.
• coord.range: If seqinfo of the tracks have seqlengths available, then use 1 to seqlength as coordinate range. If not, try to find based on ranges of features (i.e. to cover all features on that seqlevel).

An example using these arguments:

set.seed(6)
## Warning in set.seed(6): '.Random.seed' is not an integer vector but of type
## 'NULL', so ignored
pintrack <- TnT::PinTrack(GRanges("chr7", IRanges(start = sample(26300000:27000000, 4), width = 1)),
value = c(1,3,2,4), color = c("blue", "yellow", "green", "red"))
TnT::TnTGenome(
list(pintrack, btrack2),
view.range = GRanges("chr7", IRanges(26550000, 26600000)),
coord.range = IRanges(26350000, 27050000),
zoom.allow = IRanges(50000, 200000)
)