chimeraviz
2017-10-15
Contents
1 Introduction
This is the vignette of chimeraviz, an R package that automates the creation of chimeric RNA visualizations. This vignette will take you through the functionality of chimeraviz.
2 Basic features
chimeraviz allows you to import data from nine different fusion-finders: deFuse, EricScript, InFusion, JAFFA, FusionCatcher, FusionMap, PRADA, SOAPFuse, and STAR-FUSION. Getting started is easy:
# Load chimeraviz
library(chimeraviz)
# Get reference to results file from deFuse
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package="chimeraviz")
# Load the results file into a list of fusion objects
fusions <- importDefuse(defuse833ke, "hg19")Import functions for the other supported fusion-finders are similarly named (for example importEriscript or importInfusion).
A list of Fusion objects, objects that represent each fusion transcript, is now available in the list fusions.
length(fusions)## [1] 17As you can see, this list has 17 fusion objects. It is straightforward to find a specific fusion event and print information about it, or about each of the partner genes.
# Find a specific fusion event
fusion <- getFusionById(fusions, 5267)
# Show information about this fusion event
fusion## [1] "Fusion object"
## [1] "id: 5267"
## [1] "Fusion tool: defuse"
## [1] "Genome version: hg19"
## [1] "Gene names: RCC1-HENMT1"
## [1] "Chromosomes: chr1-chr1"
## [1] "Strands: +,-"
## [1] "In-frame?: FALSE"# Show information about the upstream fusion partner
upstreamPartnerGene(fusion)## [1] "PartnerGene object"
## [1] "Name: RCC1"
## [1] "ensemblId: ENSG00000180198"
## [1] "Chromosome: chr1"
## [1] "Strand: +"
## [1] "Breakpoint: 28834672"# Show information about the downstream fusion partner
downstreamPartnerGene(fusion)## [1] "PartnerGene object"
## [1] "Name: HENMT1"
## [1] "ensemblId: ENSG00000162639"
## [1] "Chromosome: chr1"
## [1] "Strand: -"
## [1] "Breakpoint: 109202584"3 Plotting
3.1 Overview plot
The overview plot is a nice way to get an overview over the nominated fusions in a sample. It will produce a circular plot like this one:
In this plot, you can see the following:
- All chromosomes with cytoband information
- Fusion events as links between locations in chromosomes (with gene names, if there is enough space in the plot)
- Red links indicate intrachromosomal fusions, while blue links indicate interchromosomal fusions
- The widths of each link varies according to how many reads support the fusion event. See the
plotCircle()documentation for more detail on how this is computed
3.1.1 Building the overview plot
All we need for the overview plot is a list of fusion events. Let’s import the SOAPfuse example data included with chimeraviz:
# Load SOAPfuse data
if(!exists("soapfuse833ke"))
soapfuse833ke <- system.file(
"extdata",
"soapfuse_833ke_final.Fusion.specific.for.genes",
package = "chimeraviz")
fusions <- importSoapfuse(soapfuse833ke, "hg38", 10)With these fusion events in the fusions variable, we’re ready to plot:
plotCircle(fusions)
3.2 Fusion reads plot
The fusion reads plot is a way to visualize the reads supporting a fusion event mapped to the putative fusion sequence. Many fusion finders report a putative fusion sequence, and by mapping reads to this sequence visualize how well the fusion event is supported. The function plotFusionReads() will, given enough data, produce a plot like this one:
As seen in the plot, this fusion event is supported by 6 paired end reads.
3.2.1 Building the Fusion Reads Plot
We have to complete a few steps in order to create the fusion reads plot. We have to:
- Align reads to the fusion sequence
- Import the alignment data into a fusion object
- Create the plot
3.2.1.1 1. Aligning reads to the fusion sequence
3.2.1.1.1 Load example fusion event
First we load an example fusion event:
# Load deFuse data
if(!exists("defuse833ke"))
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package = "chimeraviz")
fusions <- importDefuse(defuse833ke, "hg19", 1)
# Choose example fusion event
fusion <- getFusionById(fusions, 5267)3.2.1.1.2 Extracting fusion reads
Some fusion finders report which reads that gave evidence of the fusion event. By utilizing this information, we can avoid mapping all reads against the fusion sequence (that can take a while). deFuse is such a fusion finder. With the script get_fusion_fastq.pl that deFuse provides, we can extract the interesting reads. For the example fusion event, we have extracted the interesting reads into the files reads_supporting_defuse_fusion_5267.1.fq and reads_supporting_defuse_fusion_5267.2.fq. These are included in chimeraviz and can be referenced like this:
fastq1 <- system.file(
"extdata",
"reads_supporting_defuse_fusion_5267.1.fq",
package = "chimeraviz")
fastq2 <- system.file(
"extdata",
"reads_supporting_defuse_fusion_5267.2.fq",
package = "chimeraviz")3.2.1.1.3 Extract the fusion junction sequence
You can create a file containing the fusion junction sequence like this:
referenceFilename <- "reference.fa"
writeFusionReference(fusion = fusion, filename = referenceFilename)This will create a fasta file with the sequence.
3.2.1.1.4 Map fusion reads to the fusion junction sequence
The goal in this step is to align the reads you have against the fusion junction sequence. There are many ways to do this. cimeraviz offers wrapper functions for both Bowtie and Rsubread:
- Bowtie is an ultrafast and memory-efficient tool for aligning sequencing reads to reference sequences (read more here). Note that both Bowtie 1 and 2 can be used.
- Rsubread is an R package that performs (among other things) read alignment (read more here).
First we have to create an index for our reference sequence, the fusion junction sequence. Then we will align the reads against the reference. The steps are similar for both Bowtie and Rsubread:
3.2.1.1.4.1 Bowtie
# First load the bowtie functions
source(system.file(
"scripts",
"bowtie.R",
package="chimeraviz"))
# Then create index
bowtieIndex(
bowtieBuildLocation = "/path/to/bowtie-build",
referenceFasta = referenceFilename)
# And align
bowtieAlign(
bowtieLocation = "/path/to/bowtie",
referenceName = referenceFilename,
fastq1 = fastq1,
fastq2 = fastq2,
outputBamFilename = "fusionAlignment")The code above first loads the bowtie wrapper functions. This is necessary because the wrapper functions are not part of the chimeraviz package, but are added as external scripts. We then create an index for the reference sequence, and align the fusion reads to the reference. The result of this will be the files fusionAlignment.bam, fusionAlignment.bam.bai, and some additional files that Bowtie creates for indexing.
Note that you can use the parameter p with bowtieAlign to tell bowtie how many threads to use. The more you have available, the faster the alignment will complete.
3.2.1.1.4.2 Rsubread
# First load the rsubread functions
source(system.file(
"scripts",
"rsubread.R",
package="chimeraviz"))
# Then create index
rsubreadIndex(referenceFasta = referenceFilename)
# And align
rsubreadAlign(
referenceName = referenceFilename,
fastq1 = fastq1,
fastq2 = fastq2,
outputBamFilename = "fusionAlignment")The code above first loads the rsubread wrapper functions. This is necessary because the wrapper functions are not part of the chimeraviz package, but are added as external scripts. We then create an index for the reference sequence, and align the fusion reads to the reference. The result of this will be the files fusionAlignment.bam, fusionAlignment.bam.bai, and some additional files that Rsubread creates for indexing and aligning.
3.2.1.1.4.3 A note on alignment performance
If you plan to align the original fastq files against the fusion junction sequence, note that the alignment might take a while. The original fastq files used to generate all chimeraviz example data were 7.3 GB each. Helping in this regard is the fact that many fusion finders report which reads supports each fusion event. deFuse does this with the script get_fusion_fastq.pl (introduced in version 0.7 of deFuse). Other fusion finders may only output which read ids supported each fusion event. In that case, chimeraviz will help the user extract specific read ids from the original fastq files using the function fetchReadsFromFastq(). The fastq files produced by get_fusion_fastq.pl and fetchReadsFromFastq() are much smaller, resulting in the alignment step only taking a few seconds.
3.2.1.1.5 Example .bam file for this example
We have included an example .bam file with chimeraviz that contains the reads supporting fusion event 5267 aligned to the fusion junction sequence. We will continue this example with that .bam file:
if(!exists("bamfile5267"))
bamfile5267 <- system.file(
"extdata",
"5267readsAligned.bam",
package="chimeraviz")3.2.1.2 2. Import the alignment data into our fusion object
Now that we have a .bam file with the fusion reads aligned to the fusion sequence, we can import it to our fusion object:
# Add bamfile of fusion reads to the fusion oject
fusion <- addFusionReadsAlignment(fusion, bamfile5267)3.2.1.3 3. Create the plot
With all that done, we are ready to plot:
plotFusionReads(fusion)
3.3 Fusion plot
The fusion plot is the main product of chimeraviz, created with the plotFusion function. It will create a plot like this one:
Or, alternatively:
This plot holds a lot of information. You can see:
- Chromosome ideograms with a red line indicating the location of the two partner genes
- Transcript information showing exons in each partner gene
- The breakpoint between the partner genes, indicated with a red link
- Above the red link you can see the number of sequencing reads that support the fusion event
- Coverage information indicating the RNA expression level
- Genome coordinates (mega basepairs from the p-telomere)
The fusion you can see above is the RCC1-HENMT1 fusion described by Andreas M. Hoff et al. in the paper Identification of Novel Fusion Genes in Testicular Germ Cell Tumors (Cancer Research, 2016).
Note that the plot reverses genes as necessary, so that the fused genes are plotted in the “correct” (5’-to-3’) relative orientation.
3.3.1 Building the fusion plot
To build the fusion plot, we need the following:
- Information about a fusion event
- A way to get transcript information
- Coverage information from a .bam file
We will go through these steps in turn.
3.3.1.1 1. Load fusion data and find a fusion event
For this example we will load data from a deFuse run. deFuse, described in the paper deFuse: An Algorithm for Gene Fusion Discovery in Tumor RNA-Seq Data (McPherson et al., PloS Comput. Biol, 2011), is a Perl program that nominates gene fusions from RNA-seq data. chimeraviz includes example data from a deFuse run, so let’s load it using the importDefuse() function:
# First find the example data
if(!exists("defuse833ke"))
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package = "chimeraviz")
# Then load the fusion events
fusions <- importDefuse(defuse833ke, "hg19", 1)We should now have a list of fusion events in the fusions variable.
Let us try to find the RCC1-HENMT1 fusion. First, we will search our list of fusion events for fusions involving the gene RCC1 using the function getFusionByGeneName():
# See if we can find any fusions involving RCC1
getFusionByGeneName(fusions, "RCC1")## [[1]]
## [1] "Fusion object"
## [1] "id: 5267"
## [1] "Fusion tool: defuse"
## [1] "Genome version: hg19"
## [1] "Gene names: RCC1-HENMT1"
## [1] "Chromosomes: chr1-chr1"
## [1] "Strands: +,-"
## [1] "In-frame?: FALSE"As you can see, we found one fusion event involving this gene. Let us use the getFusionById() function to retrieve the fusion event:
# Extract the specific fusion
fusion <- getFusionById(fusions, 5267)We now have the fusion event we want to plot in the variable fusion.
3.3.1.2 2. Create an ensembldb object
To get transcript information we will use functions from the ensembldb package. We will use an EnsDb object to retrieve transcript information for each of the partner genes in our fusion event. Let us load an example EnsDb object:
# First find our example EnsDb file
if(!exists("edbSqliteFile"))
edbSqliteFile <- system.file(
"extdata",
"Homo_sapiens.GRCh37.74.sqlite",
package="chimeraviz")
# Then load it
edb <- ensembldb::EnsDb(edbSqliteFile)This example object, loaded from the file Homo_sapiens.GRCh37.74.sqlite that is distributed with chimeraviz, only holds transcript data for our specific example fusion transcript. If you want to, you can create a full EnsDb by downloading Ensembl data and creating an EnsDb object like this:
# Create EnsDb from a downloaded .gtf file
edbSqliteFile <- ensDbFromGtf(gtf = "Homo_sapiens.GRCh37.74.gtf")
# The function above create a .sqlite file, like the one that is included with
# chimeraviz. The path to the file is stored in the edbSqliteFile variable. To
# load the database, do this:
edb <- ensembldb::EnsDb(edbSqliteFile)If you choose to do this, note that it might take some time. You only have to do it once, though. The .sqlite file that ensDbFromGtf creates can be used in later sessions, like this:
# Create an edb object directly from the .sqlite file
edb <- ensembldb::EnsDb("Homo_sapiens.GRCh37.74.sqlite")3.3.1.3 3. Coverage information from a .bam file
We need a .bam file if we want to include coverage information in the fusion plot. chimeraviz includes an example .bam file that contains only the reads in the region of RCC1 and HENMT1. We only need a variable pointing to it:
if(!exists("fusion5267and11759reads"))
fusion5267and11759reads <- system.file(
"extdata",
"fusion5267and11759reads.bam",
package = "chimeraviz")3.3.2 Plotting the example fusion event
With the fusion event, transcript database and bamfile ready, we can create the plot with the function plotFusion():
plotFusion(
fusion = fusion,
bamfile = fusion5267and11759reads,
edb = edb,
nonUCSC = TRUE)
To collapse all transcripts into one, use the reduceTranscripts parameter:
plotFusion(
fusion = fusion,
bamfile = bamfile5267,
edb = edb,
nonUCSC = TRUE,
reduceTranscripts = TRUE)
Note the nonUCSC parameter we have set to TRUE. If the chromosome names in your .bam file are called “1” instead of “chr1”, then you need to set nonUCSC = TRUE when using plotFusion().
3.4 Fusion transcripts plot
If you are only interested in the transcripts of each partner gene in a fusion event, then it is not necessary to show all the information that is in the fusion plot. The transcripts plot is designed to give a simple view on the transcripts that might be included in the fusion transcript. It can be created like this:
# Load deFuse data
if(!exists("defuse833ke"))
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package = "chimeraviz")
fusions <- importDefuse(defuse833ke, "hg19", 1)
# Choose a fusion object
fusion <- getFusionById(fusions, 5267)
# Load edb
if(!exists("edbSqliteFile"))
edbSqliteFile <- system.file(
"extdata",
"Homo_sapiens.GRCh37.74.sqlite",
package="chimeraviz")
edb <- ensembldb::EnsDb(edbSqliteFile)
# Plot!
plotTranscripts(
fusion = fusion,
edb = edb)
The transcripts of the upstream gene is shown on top, and the transcripts of the downstream gene is shown on the bottom of the figure. As before, the exons that might be part of the fusion transcript are in darker colors. The interesting exons are also highlighted with grey boxes. Below each transcript, the gene name and the Ensembl transcript id is shown. This transcripts plot can also be shown with coverage data, if you are interested in seeing expression levels:
# Choose a fusion object
fusion <- getFusionById(fusions, 5267)
# Load edb
if(!exists("edbSqliteFile"))
edbSqliteFile <- system.file(
"extdata",
"Homo_sapiens.GRCh37.74.sqlite",
package="chimeraviz")
edb <- ensembldb::EnsDb(edbSqliteFile)
# Get reference to the .BAM file
if(!exists("fusion5267and11759reads"))
fusion5267and11759reads <- system.file(
"extdata",
"fusion5267and11759reads.bam",
package="chimeraviz")
# Plot!
plotTranscripts(
fusion = fusion,
edb = edb,
bamfile = fusion5267and11759reads,
nonUCSC = TRUE)
Similar to the fusion plot, it is also possible to reduce the transcripts for both genes into a single transcript track. Note the use of the parameter ylim to set a fixed limit for both y-axes in the plot:
# Load deFuse data
if(!exists("defuse833ke"))
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package="chimeraviz")
fusions <- importDefuse(defuse833ke, "hg19", 1)
# Choose a fusion object
fusion <- getFusionById(fusions, 5267)
# Load edb
if(!exists("edbSqliteFile"))
edbSqliteFile <- system.file(
"extdata",
"Homo_sapiens.GRCh37.74.sqlite",
package="chimeraviz")
edb <- ensembldb::EnsDb(edbSqliteFile)
# Get reference to the .BAM file
if(!exists("fusion5267and11759reads"))
fusion5267and11759reads <- system.file(
"extdata",
"fusion5267and11759reads.bam",
package="chimeraviz")
# Plot!
plotTranscripts(
fusion = fusion,
edb = edb,
bamfile = fusion5267and11759reads,
nonUCSC = TRUE,
reduceTranscripts = TRUE,
ylim = c(0,1000))
3.5 Fusion transcript plot
The fusion transcript plot shows the reduced version of all exons that could be part of a fusion transcript. This is a way to view all the possible parts of a fusion transcript merged into one.
# Load deFuse data
if(!exists("defuse833ke"))
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package="chimeraviz")
fusions <- importDefuse(defuse833ke, "hg19", 1)
# Choose a fusion object
fusion <- getFusionById(fusions, 5267)
# Load edb
if(!exists("edbSqliteFile"))
edbSqliteFile <- system.file(
"extdata",
"Homo_sapiens.GRCh37.74.sqlite",
package="chimeraviz")
edb <- ensembldb::EnsDb(edbSqliteFile)
# Plot!
plotFusionTranscript(
fusion,
edb)
As with previous visualizations, coverage data can be added.
# Load deFuse data
if(!exists("defuse833ke"))
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package="chimeraviz")
fusions <- importDefuse(defuse833ke, "hg19", 1)
# Choose a fusion object
fusion <- getFusionById(fusions, 5267)
# Load edb
if(!exists("edbSqliteFile"))
edbSqliteFile <- system.file(
"extdata",
"Homo_sapiens.GRCh37.74.sqlite",
package="chimeraviz")
edb <- ensembldb::EnsDb(edbSqliteFile)
# Get reference to the .BAM file
if(!exists("fusion5267and11759reads"))
fusion5267and11759reads <- system.file(
"extdata",
"fusion5267and11759reads.bam",
package="chimeraviz")
# Plot!
plotFusionTranscript(
fusion,
edb,
fusion5267and11759reads)
The fusion transcript plot merges the available transcripts of both genes, remove the parts that are not possible parts of the fusion transcript, and joins the included parts into a new transcript. With the coverage data included, it is easy to see the expression levels of the parts that might be included in a fusion transcript.
3.6 Fusion transcript graph plot
All that is known about a fusion event is that a fusion-finder has scored a possible link between two genes. If there are four transcript variants of the upstream gene partner and four transcript variants of the downstream gene partner, then there are in total sixteen different splice variants of the finally processed fusion transcript. And that is only if we count the known, annotated variants of each gene. How can we make sense of all this? Plotting the transcript together as in previous plots helps, but there is a better way to visualize the putative fusion transcript: As a graph. By representing transcripts as a graph, with exons as nodes and splice junctions as edges, it is much easier to get a view on the putative fusion transcript.
# Load deFuse data
if(!exists("defuse833ke"))
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package="chimeraviz")
fusions <- importDefuse(defuse833ke, "hg19", 1)
# Choose a fusion object
fusion <- getFusionById(fusions, 5267)
# Plot!
plotFusionTranscriptsGraph(
fusion,
edb)
As in previous plots, exons from the upstream gene are shown in blue, and exons from the downstream gene are shown in green. This graph plot is a compact view of all possible fusion transcripts based on the known exon boundary transcripts. It is easy to see that there are four possible versions of the first exon in the upstream gene. In the downstream gene, all transcripts first use the same six exons, but they differ in the end. In total, sixteen different transcript variants for the fusion transcript are shown in the graph.
4 Reporting and filtering
4.1 Working with Fusion objects
There are nine different functions that let you import fusion data, one for each fusion finder:
importDefuse()importEricscript()importFusioncatcher()importFusionmap()importInfusion()importJaffa()importPrada()importSoapfuse()importStarfusion()
Let us continue working with the deFuse example data included with chimeraviz:
# Get reference to results file from deFuse
if(!exists("defuse833ke"))
defuse833ke <- system.file(
"extdata",
"defuse_833ke_results.filtered.tsv",
package="chimeraviz")
# Load the results file into a list of fusion objects
fusions <- importDefuse(defuse833ke, "hg19")There are three helpful functions that allow you to manage your list of fusion objects:
# Get a specific fusion object by id
getFusionById(fusions, 5267)## [1] "Fusion object"
## [1] "id: 5267"
## [1] "Fusion tool: defuse"
## [1] "Genome version: hg19"
## [1] "Gene names: RCC1-HENMT1"
## [1] "Chromosomes: chr1-chr1"
## [1] "Strands: +,-"
## [1] "In-frame?: FALSE"# Get all fusions with a matching gene name
length(getFusionByGeneName(fusions, "RCC1"))## [1] 1# Get all fusions on a specific chromosome
length(getFusionByChromosome(fusions, "chr1"))## [1] 14.2 The fusion report
Creating a fusion report based on a list of fusion events is a nice way to get an overview on the fusions in a sample. The function createFusionReport(fusions, "output.html") will create an HTML page with an overview plot and a table of searchable, sortable fusion data. The function can be used like this:
# Load SOAPfuse data
if(!exists("soapfuse833ke"))
soapfuse833ke <- system.file(
"extdata",
"soapfuse_833ke_final.Fusion.specific.for.genes",
package = "chimeraviz")
fusions <- importSoapfuse(soapfuse833ke, "hg38", 10)
# Create report!
createFusionReport(fusions, "output.html")The result will be a file, output.html, that will look somewhat like this:
5 More information / where to get help
The first place to look for help is in this vignette or in the reference manual for chimeraviz. If the documentation and examples there are insufficient, a good place to ask for help is on the Bioconductor support site. The Biostars site may also be of help.
Any bug reports or feature ideas for chimeraviz are very welcome! Please report them as issues on Github.
6 Session Information
All code in this vignette was executed in the following environment:
sessionInfo()## R version 3.4.2 (2017-09-28)
## Platform: x86_64-apple-darwin15.6.0 (64-bit)
## Running under: OS X El Capitan 10.11.6
##
## Matrix products: default
## BLAS: /Library/Frameworks/R.framework/Versions/3.4/Resources/lib/libRblas.0.dylib
## LAPACK: /Library/Frameworks/R.framework/Versions/3.4/Resources/lib/libRlapack.dylib
##
## locale:
## [1] C/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8
##
## attached base packages:
## [1] grid stats4 parallel stats graphics grDevices utils
## [8] datasets methods base
##
## other attached packages:
## [1] chimeraviz_1.0.4 ensembldb_2.0.4 AnnotationFilter_1.0.0
## [4] GenomicFeatures_1.28.5 AnnotationDbi_1.38.2 Biobase_2.36.2
## [7] Gviz_1.20.0 GenomicRanges_1.28.6 GenomeInfoDb_1.12.3
## [10] Biostrings_2.44.2 XVector_0.16.0 IRanges_2.10.5
## [13] S4Vectors_0.14.7 BiocGenerics_0.22.1 BiocStyle_2.4.1
##
## loaded via a namespace (and not attached):
## [1] ProtGenerics_1.8.0 bitops_1.0-6
## [3] matrixStats_0.52.2 bit64_0.9-7
## [5] RColorBrewer_1.1-2 httr_1.3.1
## [7] rprojroot_1.2 Rgraphviz_2.20.0
## [9] tools_3.4.2 backports_1.1.1
## [11] R6_2.2.2 DT_0.2
## [13] rpart_4.1-11 Hmisc_4.0-3
## [15] DBI_0.7 lazyeval_0.2.0
## [17] colorspace_1.3-2 nnet_7.3-12
## [19] gridExtra_2.3 bit_1.1-12
## [21] curl_3.0 compiler_3.4.2
## [23] graph_1.54.0 htmlTable_1.9
## [25] DelayedArray_0.2.7 rtracklayer_1.36.6
## [27] bookdown_0.5 scales_0.5.0
## [29] checkmate_1.8.4 readr_1.1.1
## [31] RCircos_1.2.0 stringr_1.2.0
## [33] digest_0.6.12 Rsamtools_1.28.0
## [35] foreign_0.8-69 rmarkdown_1.6
## [37] base64enc_0.1-3 dichromat_2.0-0
## [39] pkgconfig_2.0.1 htmltools_0.3.6
## [41] BSgenome_1.44.2 htmlwidgets_0.9
## [43] rlang_0.1.2 RSQLite_2.0
## [45] BiocInstaller_1.26.1 shiny_1.0.5
## [47] jsonlite_1.5 BiocParallel_1.10.1
## [49] acepack_1.4.1 VariantAnnotation_1.22.3
## [51] RCurl_1.95-4.8 magrittr_1.5
## [53] GenomeInfoDbData_0.99.0 Formula_1.2-2
## [55] Matrix_1.2-11 Rcpp_0.12.13
## [57] munsell_0.4.3 stringi_1.1.5
## [59] yaml_2.1.14 SummarizedExperiment_1.6.5
## [61] zlibbioc_1.22.0 org.Hs.eg.db_3.4.1
## [63] plyr_1.8.4 AnnotationHub_2.8.2
## [65] blob_1.1.0 lattice_0.20-35
## [67] splines_3.4.2 hms_0.3
## [69] knitr_1.17 biomaRt_2.32.1
## [71] XML_3.98-1.9 evaluate_0.10.1
## [73] biovizBase_1.24.0 latticeExtra_0.6-28
## [75] data.table_1.10.4-2 httpuv_1.3.5
## [77] gtable_0.2.0 ggplot2_2.2.1
## [79] mime_0.5 xtable_1.8-2
## [81] ArgumentCheck_0.10.2 survival_2.41-3
## [83] tibble_1.3.4 GenomicAlignments_1.12.2
## [85] memoise_1.1.0 cluster_2.0.6
## [87] interactiveDisplayBase_1.14.0