scholarly journals Transcriptional kinetics of X-chromosome upregulation

2019 ◽  
Author(s):  
Anton J. M. Larsson ◽  
Christos Coucoravas ◽  
Rickard Sandberg ◽  
Björn Reinius
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
X Chromosome ◽  
Single Cell Rna Sequencing ◽  
Allele Specific ◽  
Mouse Cells ◽  
Kinetics Of

Ohno’s hypothesis postulates that X-chromosome upregulation rectifies X-dose imbalance relative to autosomal genes, present in two active copies per cell. Here we dissected X-upregulation into kinetics of transcription, inferred from allele-specific single-cell RNA-sequencing (scRNAseq) data from somatic mouse cells. We confirmed increased X-chromosome expression, and remarkably found that the X-chromosome achieved upregulation by elevated burst frequencies. This provides mechanistic insights into X-chromosome upregulation.

scholarly journals Estimating the Allele-Specific Expression of SNVs From 10× Genomics Single-Cell RNA-Sequencing Data

Genes ◽  
2020 ◽  
Vol 11 (3) ◽  
pp. 240 ◽  
Author(s):  
Prashant N. M. ◽  
Hongyu Liu ◽  
Pavlos Bousounis ◽  
Liam Spurr ◽  
Nawaf Alomran ◽  
...  
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Single Cells ◽  
Sequencing Data ◽  
Specific Expression ◽  
Single Nucleotide ◽  
Healthy Donors ◽  
Allele Expression ◽  
Single Cell Rna Sequencing ◽  
Allele Specific

With the recent advances in single-cell RNA-sequencing (scRNA-seq) technologies, the estimation of allele expression from single cells is becoming increasingly reliable. Allele expression is both quantitative and dynamic and is an essential component of the genomic interactome. Here, we systematically estimate the allele expression from heterozygous single nucleotide variant (SNV) loci using scRNA-seq data generated on the 10×Genomics Chromium platform. We analyzed 26,640 human adipose-derived mesenchymal stem cells (from three healthy donors), sequenced to an average of 150K sequencing reads per cell (more than 4 billion scRNA-seq reads in total). High-quality SNV calls assessed in our study contained approximately 15% exonic and >50% intronic loci. To analyze the allele expression, we estimated the expressed variant allele fraction (VAFRNA) from SNV-aware alignments and analyzed its variance and distribution (mono- and bi-allelic) at different minimum sequencing read thresholds. Our analysis shows that when assessing positions covered by a minimum of three unique sequencing reads, over 50% of the heterozygous SNVs show bi-allelic expression, while at a threshold of 10 reads, nearly 90% of the SNVs are bi-allelic. In addition, our analysis demonstrates the feasibility of scVAFRNA estimation from current scRNA-seq datasets and shows that the 3′-based library generation protocol of 10×Genomics scRNA-seq data can be informative in SNV-based studies, including analyses of transcriptional kinetics.

scholarly journals Allele-specific single-cell RNA sequencing reveals different architectures of intrinsic and extrinsic gene expression noises

2019 ◽  
Author(s):  
Mengyi Sun ◽  
Jianzhi Zhang
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Rna Sequencing ◽  
Intrinsic Noise ◽  
Mouse Fibroblast ◽  
Extrinsic Noise ◽  
Gene Expression Noise ◽  
Cell State ◽  
Single Cell Rna Sequencing ◽  
Allele Specific

ABSTRACTGene expression noise refers to the variation of the expression level of a gene among isogenic cells in the same environment, and has two sources: extrinsic noise arising from the disparity of the cell state and intrinsic noise arising from the stochastic process of gene expression in the same cell state. Due to the low throughput of the existing method for measuring the two noise components, the architectures of intrinsic and extrinsic expression noises remain elusive. Using allele-specific single-cell RNA sequencing, we here estimate the two noise components of 3975 genes in mouse fibroblast cells. Our analyses verify predicted influences of several factors such as the TATA-box and microRNA targeting on intrinsic and extrinsic noises and reveal gene function-associated noise trends implicating the action of natural selection. These findings unravel differential regulations, optimizations, and biological consequences of intrinsic and extrinsic noises and can aid the construction of desired synthetic circuits.

scholarly journals Single-cell RNA-sequencing reveals pre-meiotic X-chromosome dosage compensation in Drosophila testis

2021 ◽  
Author(s):  
Evan Witt ◽  
Zhantao Shao ◽  
Chun Hu ◽  
Henry M. Krause ◽  
Li Zhao
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
X Chromosome ◽  
Dosage Compensation ◽  
Expression Patterns ◽  
Single Cell Rna Sequencing ◽  
Germline Expression ◽  
Drosophila Testis ◽  
Male Specific ◽  
Cell Expression

AbstractDosage compensation (DC) is a mechanism by which X chromosome transcription is equalized in the somatic cells of both males and females. In male fruit flies, expression levels of the X-chromosome are increased two-fold to compensate for their single X chromosome. In testis, dosage compensation is thought to cease during meiosis, however, the timing and degree of the resulting transcriptional suppression is difficult to separate from global meiotic downregulation of each chromosome. To address this, we analyzed testis single-cell RNA-sequencing (scRNA-seq) data from two Drosophila melanogaster strains. We found evidence that the X chromosome is equally transcriptionally active as autosomes in somatic and pre-meiotic cells, and less transcriptionally active than autosomes in meiotic and post-meiotic cells. In cells experiencing dosage compensation, close proximity to MSL (male-specific lethal) chromatin entry sites (CES) correlates with increased X chromosome transcription. We found low or undetectable level of germline expression of most msl genes, mle, roX1 and roX2 via sequencing or RNA-FISH, and no evidence of germline nuclear roX1/2 localization. Our results suggest that, although DC occurs in somatic and premeiotic germ cells in Drosophila testis, there might be non-canonical factors involved in the dosage compensation. The single-cell expression patterns and enrichment statistics of detected genes can be explored interactively in our database: https://zhao.labapps.rockefeller.edu/gene-expr/.

scholarly journals Single-cell RNA-sequencing reveals pre-meiotic X-chromosome dosage compensation in Drosophila testis

PLoS Genetics ◽  
2021 ◽  
Vol 17 (8) ◽  
pp. e1009728
Author(s):  
Evan Witt ◽  
Zhantao Shao ◽  
Chun Hu ◽  
Henry M. Krause ◽  
Li Zhao
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
X Chromosome ◽  
Dosage Compensation ◽  
Expression Patterns ◽  
Single Cell Rna Sequencing ◽  
Germline Expression ◽  
Drosophila Testis ◽  
Male Specific ◽  
Cell Expression

Dosage compensation equalizes X-linked expression between XY males and XX females. In male fruit flies, expression levels of the X-chromosome are increased approximately two-fold to compensate for their single X chromosome. In testis, dosage compensation is thought to cease during meiosis; however, the timing and degree of the resulting transcriptional suppression is difficult to separate from global meiotic downregulation of each chromosome. To address this, we analyzed testis single-cell RNA-sequencing (scRNA-seq) data from two Drosophila melanogaster strains. We found evidence that the X chromosome is equally transcriptionally active as autosomes in somatic and pre-meiotic cells, and less transcriptionally active than autosomes in meiotic and post-meiotic cells. In cells experiencing dosage compensation, close proximity to MSL (male-specific lethal) chromatin entry sites (CES) correlates with increased X chromosome transcription. We found low or undetectable levels of germline expression of most msl genes, mle, roX1 and roX2 via scRNA-seq and RNA-FISH, and no evidence of germline nuclear roX1/2 localization. Our results suggest that, although dosage compensation occurs in somatic and pre-meiotic germ cells in Drosophila testis, there might be non-canonical factors involved in the dosage compensation mechanism. The single-cell expression patterns and enrichment statistics of detected genes can be explored interactively in our database: https://zhao.labapps.rockefeller.edu/gene-expr/.

scholarly journals Single-cell RNA sequencing of human, macaque, and mouse testes uncovers conserved and divergent features of mammalian spermatogenesis

2020 ◽  
Author(s):  
Adrienne Niederriter Shami ◽  
Xianing Zheng ◽  
Sarah K. Munyoki ◽  
Qianyi Ma ◽  
Gabriel L. Manske ◽  
...  
Keyword(s):  
Germ Cell ◽  
Single Cell ◽  
Rna Sequencing ◽  
Species Differences ◽  
Cell Types ◽  
Rna Turnover ◽  
Differentiation Potential ◽  
Single Cell Rna Sequencing ◽  
Kinetics Of

SummarySpermatogenesis is a highly regulated process that produces sperm to transmit genetic information to the next generation. Although extensively studied in mice, our current understanding of primate spermatogenesis is limited to populations defined by state-specific markers defined from rodent data. As between-species differences have been reported in the process duration and cellular differentiation hierarchy, it remains unclear how molecular markers and cell states are conserved or have diverged from mice to man. To address this challenge, we employ single-cell RNA-sequencing to identify transcriptional signatures of major germ and somatic cell-types of the testes in human, macaque and mice. This approach reveals differences in expression throughout spermatogenesis, including the stem/progenitor pool of spermatogonia, classical markers of differentiation, potential regulators of meiosis, the kinetics of RNA turnover during spermatid differentiation, and germ cell-soma communication. These datasets provide a rich foundation for future targeted mechanistic studies of primate germ cell development and in vitro gametogenesis.

scholarly journals Modeling allele-specific gene expression by single-cell RNA sequencing

2017 ◽  
Author(s):  
Yuchao Jiang ◽  
Nancy R Zhang ◽  
Mingyao Li
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Rna Sequencing ◽  
Specific Gene ◽  
Burst Frequency ◽  
Specific Expression ◽  
Genome Wide ◽  
Single Cell Rna Sequencing ◽  
Allelic Differences ◽  
Allele Specific

AbstractAllele-specific expression is traditionally studied by bulk RNA sequencing, which measures average expression across cells. Single-cell RNA sequencing (scRNA-seq) allows the comparison of expression distribution between the two alleles of a diploid organism and thus the characterization of allele-specific bursting. We propose SCALE to analyze genome-wide allele-specific bursting, with adjustment of technical variability. SCALE detects genes exhibiting allelic differences in bursting parameters, and genes whose alleles burst non-independently. We apply SCALE to mouse blastocyst and human fibroblast cells and find that, globally, cis control in gene expression overwhelmingly manifests as differences in burst frequency.

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