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Last updated: 2020-01-22
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Knit directory: 20170327_Psen2S4Ter_RNASeq/
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| File | Version | Author | Date | Message |
|---|---|---|---|---|
| html | b096ad9 | Steve Ped | 2020-01-21 | Recompiled index |
| Rmd | 01512da | Steve Ped | 2020-01-21 | Added initial DE analysis to index |
| Rmd | fbb6242 | Steve Ped | 2020-01-21 | Paused DE analysis |
| html | fbb6242 | Steve Ped | 2020-01-21 | Paused DE analysis |
| html | f154205 | Steve Ped | 2020-01-20 | Got rid of pointless warning |
| Rmd | bc12101 | Steve Ped | 2020-01-20 | Added bash pipeline |
| html | bc12101 | Steve Ped | 2020-01-20 | Added bash pipeline |
| html | f65bf66 | Steve Ped | 2020-01-20 | Removed License & updated text for PCA |
| Rmd | 39d0381 | Steve Ped | 2020-01-20 | Finished QC |
| html | 39d0381 | Steve Ped | 2020-01-20 | Finished QC |
| Rmd | acb4f8a | Steve Ped | 2020-01-19 | Start workflowr project. |
library(tidyverse)
library(tidygraph)
library(ggraph)samples <- read_csv("data/samples.csv") %>%
distinct(sampleName, .keep_all = TRUE) %>%
dplyr::select(sample = sampleName, sampleID, genotype) %>%
mutate(genotype = factor(genotype, levels = c("WT", "Het", "Hom")))This dataset is an analysis of RNASeq data from a 3-way comparison of WT zebrafish with Heterozygous mutants (psen2S4Ter/+) and Homozygous mutants psen2S4Ter/S4Ter. The psen2S4Ter mutant allele is expected to be a premature termination of the psen2 gene, but instead an alternate downstream start site resulted in a near full-length transcript with significant similarity to a conventional Early Onset Familial Alzhemier’s Disease mutation.
The expected comparisons are shown below:
create_ring(3) %>%
mutate(
name = paste0(levels(samples$genotype), "\n(n = 4)"),
name = factor(name, levels = name)
) %>%
activate(edges) %>%
mutate(comparison = c("Het Vs WT", "Hom Vs Het", "Hom Vs WT")) %>%
ggraph(layout = "kk") +
geom_edge_link2(
aes(label = comparison),
angle_calc = "along",
label_dodge = unit(0.02, "npc"),
start_cap = circle(0.09, "npc"),
end_cap = circle(0.09, "npc"),
label_size = 5,
arrow = arrow(
length = unit(0.06, "npc"),
ends = "both",
type = "closed"
)
) +
geom_node_label(
aes(label = name, colour = name),
size = 5,
fill = rgb(1,1,1,0.7),
label.padding = unit(0.4, "lines")
) +
scale_y_continuous(expand = expand_scale(0.1)) +
scale_x_continuous(expand = expand_scale(0.1)) +
theme_void() +
theme(
legend.position = "none"
) 
All samples were female, 6 months old and raised in the same tank as a family, to minimise variability. This age represents sexual maturity and is expected to model a pre-symptomatic brain. Fish were all killed and genotyped by tail-clipping on 07-07-2016.
RNAseq was performed on total RNA (i.e. ribo-reduced) from whole brain tissue, with \(n=4\) samples in each group. Sequencing was performed by the sequencing facility at the Centre for Cancer Biology in Adelaide, using an Illimuna NextSeq. Reads were provided in as paired-end, 150bp reads.
devtools::session_info()─ Session info ───────────────────────────────────────────────────────────────
setting value
version R version 3.6.2 (2019-12-12)
os Ubuntu 18.04.3 LTS
system x86_64, linux-gnu
ui X11
language en_AU:en
collate en_AU.UTF-8
ctype en_AU.UTF-8
tz Australia/Adelaide
date 2020-01-22
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