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RNA-seq on DGRP lines and F1 crosses

ABSTRACT: RNA-seq on DGRP lines and F1 crosses

PROVIDER: PRJEB3230 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR165901.fastq.gz	Fastqsanger.gz
	ERR165902.fastq.gz	Fastqsanger.gz
	ERR165903.fastq.gz	Fastqsanger.gz
	ERR165904.fastq.gz	Fastqsanger.gz
	ERR165905.fastq.gz	Fastqsanger.gz

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RNA-seq on DGRP lines and F1 crosses

Project description:We performed mRNA sequencing of reciprocal F1 female hybrids from two crosses (362/765 and 517/765) and the parental DGRP lines (362, 517 and 765). Specifically, we aimed to identify whether transcripts predicted to be regulated by cis-eQTLs exhibit a significant allele-specific bias in gene expression. Since both alleles act in the cross in the same trans environment, differential expression in the F1 is a direct measure of cis-regulatory activity.

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Drosophila Genetic Reference Panel (DGRP) F1 RNA-seq - a comparison to 3prime-tag-seq variance decomposition analysis

Project description:Crosses were carried out between two pairs of lines from the Drosophila Genetic Reference Panel (DGRP): Line 765 (mother) x Line 517 (father) and Line 517 (mother) x Line 362 (father). Embryos resulting from these crosses (as well as from each of the parental lines) was collected at three stages of development: 2-4h, 6-8h, and 10-12h after egg laying. Following standard procedure, three, one-hour pre-lays were collected prior to the experiment (to synchronise embryo age) and all collections were done at 25C. Collections were also made of the reciprocal crosses at 2-4h after egg laying. To validate the variance decomposiiton analysis, we focused our attention on the primary F1 crosses 765 x 517 and 517 x 362. Estimates of line-specific expression, and associated methods, can be found as supplements to Cannavo et al.

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Project description:Full-length, directional RNA-Seq data from a panel of 8 F1 hybid D. melanogaster lines along with matched RNA-Seq data from the two parental lines for one of the F1 crosses. All paternal fly lines were taken from the Drosophila Genetic Reference Panel crossed to a common mother (PMID31308546). Data were collected at three time points (2-4h, 6-8h, 10-12h at 25C) with two biological replicates per collection.

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A novel statistical approach for jointly analyzing RNA-Seq data from F1 reciprocal crosses and inbred lines.

Project description:RNA sequencing (RNA-seq) not only measures total gene expression but may also measure allele-specific gene expression in diploid individuals. RNA-seq data collected from F1 reciprocal crosses in mice can powerfully dissect strain and parent-of-origin effects on allelic imbalance of gene expression. In this article, we develop a novel statistical approach to analyze RNA-seq data from F1 and inbred strains. Method development was motivated by a study of F1 reciprocal crosses derived from highly divergent mouse strains, to which we apply the proposed method. Our method jointly models the total number of reads and the number of allele-specific reads of each gene, which significantly boosts power for detecting strain and particularly parent-of-origin effects. The method deals with the overdispersion problem commonly observed in read counts and can flexibly adjust for the effects of covariates such as sex and read depth. The X chromosome in mouse presents particular challenges. As in other mammals, X chromosome inactivation silences one of the two X chromosomes in each female cell, although the choice of which chromosome to be silenced can be highly skewed by alleles at the X-linked X-controlling element (Xce) and stochastic effects. Our model accounts for these chromosome-wide effects on an individual level, allowing proper analysis of chromosome X expression. Furthermore, we propose a genomic control procedure to properly control type I error for RNA-seq studies. A number of these methodological improvements can also be applied to RNA-seq data from other species as well as other types of next-generation sequencing data sets. Finally, we show through simulations that increasing the number of samples is more beneficial than increasing the library size for mapping both the strain and parent-of-origin effects. Unless sample recruiting is too expensive to conduct, we recommend sequencing more samples with lower coverage.

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Project description:DGRP lines were raised at 18 degree celsius, adult mated whole fly expression was estimated using microarrays. The same design was previous used to measure expression at 25 degree celsius (E-MTAB-3216).

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Project description:The extreme generalist two-spotted spider mite, Tetranychus urticae, which is documented to feed on more than 1100 plant hosts, is becoming an increasingly important agricultural pest. Historically, as studies of plant-herbivore interactions have focused largely on insects, considerably less research has investigated plant responses to spider mite herbivores, especially in grasses. To identify intraspecific differences in maize response to T. urticae, we collected RNA-seq data from three maize (Zea mays) inbred lines (B73, B75 and B49) as well as two F1 lines arising from crosses between B73 x B75 and B73 x B96. For each maize line, RNA-seq data was collected from uninfested leaves (control) and leaves infested with T. urticae for 24 hours.

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