Allele-specific Expression Variation Research Articles

The transcriptional legacy of developmental stochasticity

Genetic and environmental variation are key contributors during organism development, but the influence of minor perturbations or noise is difficult to assess. This study focuses on the stochastic variation in allele-specific expression that persists through cell divisions in the nine-banded armadillo (Dasypus novemcinctus). We investigated the blood transcriptome of five wild monozygotic quadruplets over time to explore the influence of developmental stochasticity on gene expression. We identify an enduring signal of autosomal allelic variability that distinguishes individuals within a quadruplet despite their genetic similarity. This stochastic allelic variation, akin to X-inactivation but broader, provides insight into non-genetic influences on phenotype. The presence of stochastically canalized allelic signatures represents a novel axis for characterizing organismal variability, complementing traditional approaches based on genetic and environmental factors. We also developed a model to explain the inconsistent penetrance associated with these stochastically canalized allelic expressions. By elucidating mechanisms underlying the persistence of allele-specific expression, we enhance understanding of development’s role in shaping organismal diversity.

Gene age shapes the transcriptional landscape of sexual morphogenesis in mushroom-forming fungi (Agaricomycetes).

Multicellularity has been one of the most important innovations in the history of life. The role of gene regulatory changes in driving transitions to multicellularity is being increasingly recognized; however, factors influencing gene expression patterns are poorly known in many clades. Here, we compared the developmental transcriptomes of complex multicellular fruiting bodies of eight Agaricomycetes and Cryptococcus neoformans, a closely related human pathogen with a simple morphology. In-depth analysis in Pleurotus ostreatus revealed that allele-specific expression, natural antisense transcripts, and developmental gene expression, but not RNA editing or a 'developmental hourglass,' act in concert to shape its transcriptome during fruiting body development. We found that transcriptional patterns of genes strongly depend on their evolutionary ages. Young genes showed more developmental and allele-specific expression variation, possibly because of weaker evolutionary constraint, suggestive of nonadaptive expression variance in fruiting bodies. These results prompted us to define a set of conserved genes specifically regulated only during complex morphogenesis by excluding young genes and accounting for deeply conserved ones shared with species showing simple sexual development. Analysis of the resulting gene set revealed evolutionary and functional associations with complex multicellularity, which allowed us to speculate they are involved in complex multicellular morphogenesis of mushroom fruiting bodies.

Targeted RNA-seq improves efficiency, resolution, and accuracy of allele specific expression for human term placentas.

Genomic imprinting is an epigenetic mechanism that results in allele-specific expression (ASE) based on the parent of origin. It is known to play a role in the prenatal and postnatal allocation of maternal resources in mammals. ASE detected by whole transcriptome RNA-seq (wht-RNAseq) has been widely used to analyze imprinted genes using reciprocal crosses in mice to generate large numbers of informative SNPs. Studies in humans are more challenging due to the paucity of SNPs and the poor preservation of RNA in term placentas and other tissues. Targeted RNA-seq (tar-RNAseq) can potentially mitigate these challenges by focusing sequencing resources on the regions of interest in the transcriptome. Here, we compared tar-RNAseq and wht-RNAseq in a study of ASE in known imprinted genes in placental tissue collected from a healthy human cohort in Mali, West Africa. As expected, tar-RNAseq substantially improved the coverage of SNPs. Compared to wht-RNAseq, tar-RNAseq produced on average four times more SNPs in twice as many genes per sample and read depth at the SNPs increased fourfold. In previous research on humans, discordant ASE values for SNPs of the same gene have limited the ability to accurately quantify ASE. We show that tar-RNAseq reduces this limitation as it unexpectedly increased the concordance of ASE between SNPs of the same gene, even in cases of degraded RNA. Studies aimed at discovering associations between individual variation in ASE and phenotypes in mammals and flowering plants will benefit from the improved power and accuracy of tar-RNAseq.

Pervasive Inter-Individual Variation in Allele-Specific Expression in Monozygotic Twins

Despite being developed from one zygote, heterokaryotypic monozygotic (MZ) co-twins exhibit discordant karyotypes. Epigenomic studies in biological samples from heterokaryotypic MZ co-twins are of the most significant value for assessing the effects on gene- and allele-specific expression of an extranumerary chromosomal copy or structural chromosomal disparities in otherwise nearly identical germline genetic contributions. Here, we use RNA-Seq data from existing repositories to establish within-pair correlations for the breadth and magnitude of allele-specific expression (ASE) in heterokaryotypic MZ co-twins discordant for trisomy 21 and maternal 21q inheritance, as well as homokaryotypic co-twins. We show that there is a genome-wide disparity at ASE sites between the heterokaryotypic MZ co-twins. Although most of the disparity corresponds to changes in the magnitude of biallelic imbalance, ASE sites switching from either strictly monoallelic to biallelic imbalance or the reverse occur in few genes that are known or predicted to be imprinted, subject to X-chromosome inactivation or A-to-I(G) RNA edited. We also uncovered comparable ASE differences between homokaryotypic MZ twins. The extent of ASE discordance in MZ twins (2.7%) was about 10-fold lower than the expected between pairs of unrelated, non-twin males or females. The results indicate that the observed within-pair dissimilarities in breadth and magnitude of ASE sites in the heterokaryotypic MZ co-twins could not solely be attributable to the aneuploidy and the missing allelic heritability at 21q.

Modeling allele-specific expression at the gene and SNP levels simultaneously by a Bayesian logistic mixed regression model

BackgroundHigh-throughput sequencing experiments, which can determine allele origins, have been used to assess genome-wide allele-specific expression. Despite the amount of data generated from high-throughput experiments, statistical methods are often too simplistic to understand the complexity of gene expression. Specifically, existing methods do not test allele-specific expression (ASE) of a gene as a whole and variation in ASE within a gene across exons separately and simultaneously.ResultsWe propose a generalized linear mixed model to close these gaps, incorporating variations due to genes, single nucleotide polymorphisms (SNPs), and biological replicates. To improve reliability of statistical inferences, we assign priors on each effect in the model so that information is shared across genes in the entire genome. We utilize Bayesian model selection to test the hypothesis of ASE for each gene and variations across SNPs within a gene. We apply our method to four tissue types in a bovine study to de novo detect ASE genes in the bovine genome, and uncover intriguing predictions of regulatory ASEs across gene exons and across tissue types. We compared our method to competing approaches through simulation studies that mimicked the real datasets. The R package, BLMRM, that implements our proposed algorithm, is publicly available for download at https://github.com/JingXieMIZZOU/BLMRM.ConclusionsWe will show that the proposed method exhibits improved control of the false discovery rate and improved power over existing methods when SNP variation and biological variation are present. Besides, our method also maintains low computational requirements that allows for whole genome analysis.

Abstract 1584: Transcriptome analysis links immune genes allelic expression imbalances to lung cancer

Abstract Background: Genome-wide association study (GWAS) have identified over 45 susceptibility loci for lung cancer; many studies including our own group, have focused on low-frequency and rare coding variants using fine mapping and exome sequencing. This strategy, however, has met with limited success as about 90% of GWAS hits are noncoding and act primarily through altering transcriptional regulation in an allele-specific manner. The RNA-Seq based allele-specific expression (ASE) analysis affords an innovative approach to study preferential expression of an allele in direct relationship to its genotype, providing information on cis-regulatory effects for the expression of putative genes. However currently, there are no lung cancer studies that have rigorously evaluated the ASE variation in lung tumor and adjacent tissues. Methods: Leveraging The Cancer Genome Atlas (TCGA) resource, we performed transcriptomic-wide ASE analysis using existing RNA-Seq datasets of paired tumor and adjacent tissues from 54 lung adenocarcinoma patients. We first quantified the RNA read counts of Referent and Alternate alleles of heterozygous variants, then evaluated the allelic imbalance on a per-sample basis using Beta-binomial test, and explored the differential ASE between tumor and adjacent tissues using paired Wilcoxon test. Functional regulatory consequences were generated from Ensembl Variant Effect Predictor. Results: We identified total 208 significant ASEs, including 35 tissue-specific (only in tumor or only in adjacent), 28 sharing, and 145 differential variants. Of the 208 candidates, 41 were from the human leukocyte antigen (HLA) locus (primary DQA2, DQB1, DRB1, H and J), 26 were from the immunoglobulin (IG) superfamily (primary IGH, IGL, IGK and F11R). About 80% candidates were noncoding (mostly in 5’ and 3’ untranslated regions) and with regulatory features (21 promoter, seven enhancer, seven open chromatin region, two induce nonsense-mediated mRNA decay, one CTCF-binding site, and one transcription factor binding site). Other top genes included MDM2, APOL1, and CTSB. Pathway analyses revealed 27 genes involved in immune response pathway, and 12 genes involved in HLA antigen processing and presentation pathway. Conclusion: This study is the first transcriptomics ASE analysis in lung adenocarcinoma. The key somatic cis-regulatory ASE variants identified from this study, especially immunogenic allelic variations from HLA and IG genes, could be used for identifying high-risk individuals for targeted lung cancer checkpoint blockade and related immunotherapies. Citation Format: Yanhong Liu, Spiridon Tsavachidis, Farrah Kheradmand, Margaret R. Spitz, Chris Amos. Transcriptome analysis links immune genes allelic expression imbalances to lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1584.

Allele-specific expression variation at different ploidy levels in Squalius alburnoides

Allopolyploid plants are long known to be subject to a homoeolog expression bias of varying degree. The same phenomenon was only much later suspected to occur also in animals based on studies of single selected genes in an allopolyploid vertebrate, the Iberian fish Squalius alburnoides. Consequently, this species became a good model for understanding the evolution of gene expression regulation in polyploid vertebrates. Here, we analyzed for the first time genome-wide allele-specific expression data from diploid and triploid hybrids of S. alburnoides and compared homoeolog expression profiles of adult livers and of juveniles. Co-expression of alleles from both parental genomic types was observed for the majority of genes, but with marked homoeolog expression bias, suggesting homoeolog specific reshaping of expression level patterns in hybrids. Complete silencing of one allele was also observed irrespective of ploidy level, but not transcriptome wide as previously speculated. Instead, it was found only in a restricted number of genes, particularly ones with functions related to mitochondria and ribosomes. This leads us to hypothesize that allelic silencing may be a way to overcome intergenomic gene expression interaction conflicts, and that homoeolog expression bias may be an important mechanism in the achievement of sustainable genomic interactions, mandatory to the success of allopolyploid systems, as in S. alburnoides.

Unperturbed expression bias of imprinted genes in schizophrenia

How gene expression correlates with schizophrenia across individuals is beginning to be examined through analyses of RNA-seq from postmortem brains of individuals with disease and control brains. Here we focus on variation in allele-specific expression, following up on the CommonMind Consortium (CMC) RNA-seq experiments of nearly 600 human dorsolateral prefrontal cortex (DLPFC) samples. Analyzing the extent of allelic expression bias—a hallmark of imprinting—we find that the number of imprinted human genes is consistent with lower estimates (≈0.5% of all genes), and thus contradicts much higher estimates. Moreover, the handful of putatively imprinted genes are all in close genomic proximity to known imprinted genes. Joint analysis of the imprinted genes across hundreds of individuals allowed us to establish how allelic bias depends on various factors. We find that age and genetic ancestry have gene-specific, differential effect on allelic bias. In contrast, allelic bias appears to be independent of schizophrenia.

Putative bovine topological association domains and CTCF binding motifs can reduce the search space for causative regulatory variants of complex traits

BackgroundTopological association domains (TADs) are chromosomal domains characterised by frequent internal DNA-DNA interactions. The transcription factor CTCF binds to conserved DNA sequence patterns called CTCF binding motifs to either prohibit or facilitate chromosomal interactions. TADs and CTCF binding motifs control gene expression, but they are not yet well defined in the bovine genome. In this paper, we sought to improve the annotation of bovine TADs and CTCF binding motifs, and assess whether the new annotation can reduce the search space for cis-regulatory variants.ResultsWe used genomic synteny to map TADs and CTCF binding motifs from humans, mice, dogs and macaques to the bovine genome. We found that our mapped TADs exhibited the same hallmark properties of those sourced from experimental data, such as housekeeping genes, transfer RNA genes, CTCF binding motifs, short interspersed elements, H3K4me3 and H3K27ac. We showed that runs of genes with the same pattern of allele-specific expression (ASE) (either favouring paternal or maternal allele) were often located in the same TAD or between the same conserved CTCF binding motifs. Analyses of variance showed that when averaged across all bovine tissues tested, TADs explained 14% of ASE variation (standard deviation, SD: 0.056), while CTCF explained 27% (SD: 0.078). Furthermore, we showed that the quantitative trait loci (QTLs) associated with gene expression variation (eQTLs) or ASE variation (aseQTLs), which were identified from mRNA transcripts from 141 lactating cows’ white blood and milk cells, were highly enriched at putative bovine CTCF binding motifs. The linearly-furthermost, and most-significant aseQTL and eQTL for each genic target were located within the same TAD as the gene more often than expected (Chi-Squared test P-value < 0.001).ConclusionsOur results suggest that genomic synteny can be used to functionally annotate conserved transcriptional components, and provides a tool to reduce the search space for causative regulatory variants in the bovine genome.

Association mapping reveals the role of purifying selection in the maintenance of genomic variation in gene expression

The evolutionary forces that maintain genetic variation in quantitative traits within populations remain poorly understood. One hypothesis suggests that variation is under purifying selection, resulting in an excess of low-frequency variants and a negative correlation between minor allele frequency and selection coefficients. Here, we test these predictions using the genetic loci associated with total expression variation (eQTLs) and allele-specific expression variation (aseQTLs) mapped within a single population of the plant Capsella grandiflora. In addition to finding eQTLs and aseQTLs for a large fraction of genes, we show that alleles at these loci are rarer than expected and exhibit a negative correlation between phenotypic effect size and frequency. Overall, our results show that the distribution of frequencies and effect sizes of the loci responsible for local expression variation within a single outcrossing population are consistent with the effects of purifying selection.

Abstract 2084: Genetic variation at a cis-acting C/EBPG binding site is associated with allele-specific ERCC5 transcript expression

Abstract Background: CCAAT/enhancer-binding protein gamma (C/EBPG) transcription factor expression is correlated with that of ERCC5 and other key DNA repair genes in normal bronchial epithelial cells (NBEC) suggesting a regulatory role. In prior studies, ERCC5 transcript expression was increased in a human lung carcinoma cell line H23 following CEBPG overexpression and in NBEC from 81 subjects, A allele at putative ERCC5 cis-regulatory SNP (rSNP) rs751402 and T allele at rSNP rs2296147 were associated with higher expression of ERCC5 marker SNP rs1047768 T allele transcript. rs751402 is located in open chromatin region identified by FAIRE-seq in NBEC and variation at rs751402 is predicted to alter binding of C/EBP. These studies support the hypothesis that allelic differential affinity to C/EBPG at rs751402 contributes to hereditary inter-individual variation in regulation of ERCC5 either directly or through interaction with complexes bound at rs2296147. The purpose of this study was to further investigate the role of C/EBPG in ERCC5 cis-regulation in an independent cohort of subjects and lung cancer cell lines. Methods: We knocked-down C/EBPG transcript level by C/EBPG siRNA transfection in human non-small cell lung carcinoma cell line H1703. Total and allele-specific expression (ASE) at rs1047768 was measured through multiplex competitive PCR-based amplicon sequencing library preparation followed by Illumina HiSeq next generation sequencing (NGS). This NGS controls for inter-target variation in PCR amplification during library preparation by measuring each transcript native template relative to a known number of synthetic competitive template internal standard copies. The genotype at rs751402 and rs2296147 in NBEC from 78 subjects and 14 human lung carcinoma cell lines was determined by TaqMan SNP genotyping assays. Direct assessment of the syntenic relationship of alleles in gDNA from poly-heterozygous individuals was assessed by allele-specific PCR followed by sequencing. Results: CEBPG transcript expression was knocked-down by 93% in H1703 cells and this was associated with 4-fold reduction in the ERCC5 transcript level at rs1047768. ERCC5 displayed significant inter-individual variation in allele specific expression (ASE) in NBEC from 85 subjects. Thirty nine out of 92 subjects including 3 cell lines were heterozygous at rs751402 and rs2296147 rSNPs and rs1047768 marker SNP and will be assessed for haplotypes comprising those sites. Conclusions: Results are consistent with CEBPG regulation of ERCC5 in the cell line H1703. The results obtained will enable us to test the hypothesis that haplotypes comprising particular alleles syntenic between rs1047768 and rs751402 are associated with higher allele-specific ERCC5 transcript abundance. Cell lines heterozygous at three sites will be subjected to CEBPG up and/or down regulation to assess effect on allele-specific ERCC5 expression at rs1047768. Citation Format: Xiaolu Zhang, Jiyoun Yeo, Erin Crawford, James C. Willey. Genetic variation at a cis-acting C/EBPG binding site is associated with allele-specific ERCC5 transcript expression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2084. doi:10.1158/1538-7445.AM2015-2084

Abstract 4156: Inter-individual variation in allele specific expression of catalase (CAT) in normal bronchial epithelial cells and association of putative cis-regulatory CAT SNP rs12807961 with lung cancer risk

Abstract Background: Catalase (CAT) is a key antioxidant gene expressed at high levels in most human tissues, including normal bronchial epithelial cells (NBEC). NBEC CAT expression is more disperse (more high and low extreme values) among subjects with cancer compared to controls. CAT shares this property with 14 other antioxidant and DNA repair genes comprised by the Lung Cancer Risk Test (LCRT) reported from this laboratory. We hypothesize that inter-individual variation in CAT regulation in NBEC is in part due to inter-individual variation at one or more cis-regulatory single nucleotide polymorphisms (SNPs). If so, this should manifest as inter-individual variation in allele-specific CAT expression in NBEC. Through funding in part from RC2 CA148572 and HL108016 we collected NBEC samples from over 500 subjects at risk for lung cancer. In this pilot study, we assessed allele-specific and total expression of multiple genes in NBEC samples from 85 subjects and assessed the genotype at putative cis-regulatory CAT SNP rs12807961 in gDNA from 95 subjects. Methods: RNA extracted from normal bronchial airway brush specimens of 85 subjects (26 cancer cases and 59 non-cancer controls) was reverse transcribed. Using next generation sequencing (NGS), allele-specific expression (ASE) was measured as allelic imbalance in each cDNA at three marker SNPs in the CAT coding region (rs1049982, rs769217, and rs704724) and one putative regulatory SNP (rs12807961) that was 4364 bases upstream of transcription start site, using gDNA as control. Specifically, each cDNA and matched peripheral blood cell gDNA sample was subjected to targeted competitive template multiplex PCR amplicon library generation followed by NGS (Blomquist et al, PLOS one, 2013) on Illumina Hiseq platform. The genotype at putative cis-regulatory SNP rs12807961 was assessed in gDNA from 95 subjects including those assessed for ASE (a total of 31 cancer cases and 64 non-cancer controls) using a TaqMan® SNP Genotyping Assay. Results: Among heterozygotes, there was significant inter-individual variation in cDNA allelic imbalance at rs1049982 (p&amp;lt;0.001, n=40) and rs769217 (p&amp;lt;0.001, n=28) as measured by F-test using matched gDNA controls. In this cohort there was insufficient number of heterozygotes at rs704724 (n=2) to assess ASE. Among all 95 subjects assessed for rs12807961 genotype, nine were homozygous minor allele at the rs12807961 CAT SNP. Of these, 7/31 cancer cases and 2/64 non-cancer controls were homozygous minor allele. This difference was significant by two-tailed Fisher exact test (P&amp;lt;0.05) following Bonferroni adjustment for multiple testing. Conclusions: These data support the hypothesis that cis-regulatory DNA variants contribute to inter-individual variation in CAT regulation in NBEC and that this is associated with lung cancer risk. Citation Format: Jiyoun Yeo, Xaiolu Zhang, Erin L. Crawford, James C. Willey. Inter-individual variation in allele specific expression of catalase (CAT) in normal bronchial epithelial cells and association of putative cis-regulatory CAT SNP rs12807961 with lung cancer risk. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4156. doi:10.1158/1538-7445.AM2014-4156

Genotype–phenotype mapping in a post-GWAS world

Understanding how metabolic reactions, cell signaling, and developmental pathways translate the genome of an organism into its phenotype is a grand challenge in biology. Genome-wide association studies (GWAS) statistically connect genotypes to phenotypes, without any recourse to known molecular interactions, whereas a molecular biology approach directly ties gene function to phenotype through gene regulatory networks (GRNs). Using natural variation in allele-specific expression, GWAS and GRN approaches can be merged into a single framework via structural equation modeling (SEM). This approach leverages the myriad of polymorphisms in natural populations to elucidate and quantitate the molecular pathways that underlie phenotypic variation. The SEM framework can be used to quantitate a GRN, evaluate its consistency across environments or sexes, identify the differences in GRNs between species, and annotate GRNs de novo in non-model organisms.

Abstract 4988: Cis-acting genetic variation is associated with altered allele-specific expression of CEBPG transcript in normal human bronchial epithelium

Abstract CEBPG (CCAAT/enhancer binding protein gamma) is one of six known members of the CEBP family of bZIP transcription factors. Gene targeted knock-out of CEBPG transcription factor in mice results in early perinatal death, presumably due to observed emphysematous lung histology and abnormal development of alveolar pneumocytes (Kaisho et al, 1999). In recent studies, we observed that: 1) CEBPG transcript expression in normal human bronchial epithelial cells (NBEC) varies greater than 20-fold among individuals (Mullins et al, 2005), 2) the pattern of CEBPG transcript expression in NBEC is different between lung cancer cases and controls (Blomquist et al, 2009), and 3) transcript expression of a key DNA repair gene, ERCC5 (XPG), is partly regulated by CEBPG in NBEC (Crawford et al, 2007). In this study we hypothesized that variation in CEBPG transcript expression patterns in NBEC may, in part, be due to cis-acting genetic variation. In order to test this hypothesis, Allele-Specific Expression (ASE) was measured at transcribed polymorphic site rs2772 in Exon 2 of CEBPG in NBEC cDNA of fifteen individuals heterozygous at rs2772 using allele-specific competitive PCR. ASE at rs2772 was then assessed for association with genotype at the seven polymorphic sites (rs736682, rs17530479, rs17530508, rs1469084, rs16968029, rs3745968, rs36101103) representing the most common haplotypes in the promoter, intron and 3′-untranslated regions of CEBPG gene. CEBPG rs2772 A allele was expressed, on average, 2.34 fold greater than C allele (p &amp;lt; 0.001) in 20% (3 out of 15) of NBEC cDNA samples, among individuals heterozygous at both rs2772 and rs17530479 in the 3′-untranslated and promoter regions of CEBPG, respectively. These findings are consistent with previous reports that in 20% of samples, CEBPG rs2772 A allele was expressed 2.42 fold greater than C allele (Lo et al, 2003). We also observed that 3 out of 9 NBEC samples obtained from individuals diagnosed with lung cancer exhibited rs2772 A:C allelic ratios significantly higher than genomic DNA controls, while 0 out 6 samples from individuals without lung cancer exhibited altered ASE. Of note, rs2772 and rs17530479 polymorphic sites are in linkage disequilibrium with rs10518275 polymorphic site, which was recently associated with severity of cystic fibrosis lung disease (Gu et al, 2009). Here we demonstrate that cis-acting variation in linkage with rs2772 and rs17530479 polymorphic sites is associated with variation in ASE of CEBPG and lung cancer diagnosis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4988.

Dynamic variation in allele-specific gene expression of Paraoxonase-1 in murine and human tissues

Differential allelic expression has been shown to be common in mice, humans and maize, and variability in the expression of polymorphic alleles has been associated with human disease. Here, we describe the differential expression pattern of Paraoxonase-1, a gene involved in lipid metabolism and implicated in the formation of atherosclerotic lesions. We measured the expression of the murine Paraoxonase-1 gene (Pon1) in livers at different stages of embryonic development using F1 hybrid crosses and quantified the transcriptional level of both parental alleles. Using human foetal tissues, we analysed the expression of the human orthologue (PON1) and found monoallelic or preferential allelic expression in 6/7 and 4/4 samples from liver and pancreas, respectively. We observed that Pon1 does not show a parent-of-origin preference in its allelic expression, but has dramatic variations in allele-specific expression occurring throughout development. This study has important repercussions in the analysis of haplotypes at disease loci, since it implies that the expression of polymorphic alleles can be unequal and dynamic.

Allelic variation of gene expression in maize hybrids.

Allelic expression variation of nonimprinted autosomal genes has recently been uncovered in mouse hybrids and humans. The allelic expression variation is attributed to differences in noncoding DNA sequences and does not involve epigenetic regulation or gene imprinting. This expression variation is suggested to play important roles in determining phenotypic diversity. Virtually nothing is known about such allele-specific expression variation in a hybrid plant where two alleles are compared in the same genetic context. We examined parental transcript accumulation in maize (Zea mays) hybrids using allele-specific RT-PCR analysis. Among 15 genes analyzed, 11 showed differences at the RNA level, ranging from unequal expression of the two alleles (biallelic) to expression of a single allele (monoallelic). Maternal or paternal transmission had little effect on the allele-specific transcript ratio of nearly all genes analyzed, suggesting that parent-of-origin effect was minimal. We analyzed the allelic difference in genetically contrasting hybrids and hybrids under high planting density and drought stress. Whereas a genetically improved modern hybrid expressed both alleles, a less improved old hybrid frequently showed mono-allelic expression. Furthermore, the two alleles in the hybrid responded differentially to abiotic stresses. The results of allele-specific regulation in different tissues in responding to environment and stress suggest an unequivalent function of the parental alleles in the hybrid, which may have an impact on heterosis.