High temperature increases centromere-mediated genome elimination frequency and enhances haploid induction in Arabidopsi
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ABSTRACT:
PROVIDER: PRJEB58839 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR10761813_1.fastq.gz | Fastqsanger.gz | |||
| ERR10761813_2.fastq.gz | Fastqsanger.gz | |||
| ERR10761814_1.fastq.gz | Fastqsanger.gz | |||
| ERR10761814_2.fastq.gz | Fastqsanger.gz | |||
| ERR10761815_1.fastq.gz | Fastqsanger.gz |
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Project description:Double haploid production is the most effective way to create true-breeding lines in a single generation. In Arabidopsis, haploid induction via mutation of the centromere-specific histone H3 (cenH3) has been shown when the mutant is outcrossed to the wild-type, and the wild-type genome remains in the haploid progeny. However, factors that affect haploid induction are still poorly understood. Here, we report that a mutant of the cenH3 assembly factor Kinetochore Null2 (KNL2) can be used as a haploid inducer when pollinated by the wild-type. We discovered that short-term temperature stress of the knl2 mutant increased the efficiency of haploid induction 10-fold. We also demonstrated that a point mutation in the CENPC-k motif of KNL2 is sufficient to generate haploid-inducing lines, suggesting that haploid-inducing lines in crops can be identified in a naturally occurring or chemically induced mutant population, avoiding the generic modification (GM) approach at any stage. Furthermore, a cenh3-4 mutant functioned as a haploid inducer in response to short-term heat stress, even though it did not induce haploids under standard conditions. Thus, we identified KNL2 as a new target gene for the generation of haploid-inducer lines and showed that exposure of centromeric protein mutants to high temperature strongly increases their haploid induction efficiency.
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Project description:Simple and consistent production of haploid is always an appealing pursuit for both crop breeders and researchers. Although diverse strategies have been developed to produce haploids over the past decades, most of them are applicable in only a limited number of plant species. In 2010, Ravi and Chan reported that haploid Arabidopsis thaliana plants can be efficiently induced through the introduction of a single genetic alteration in centromere histone H3 (CENH3). Subsequent studies demonstrated that haploids can be efficiently induced either through genetic engineering of CENH3 N-terminal tail or histone fold domain or by replacing CENH3 with an ortholog. The mutation of a pollen-specific phospholipase gene, MATRILINEAL (MTL) has been revealed to trigger the haploid induction (HI) in maize, which present another promising HI approach by the editing of MTL in plant. Here, we review the progress of the CENH3-medialed HI and propose a revised centromere-size model by suggesting a competitive loading process between wild-type and mutant CENH3 during HI. This model can explain both the findings of HI failure when wild-type and mutant CENH3 genes are coexpressed and the alien centromere loading of CENH3 in stable hybrids. In addition, we review the current understanding of MTL-mediated HI in plant. The conservation of CENH3 and MTL in plants indicates wide potential application for HI. We discuss the utility and potential of these two methods in crops by comparing their mechanisms and applications to date in plants. This review will promote the study and application of both CENH3- and MTL-mediated haploid induction in plants.
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Project description:Recent success in the derivation of mouse haploid embryonic stem cells from androgenetic blastocysts (ahESCs) has provided new avenues for the generation of genetically modified animals. However, the efficiency to produce viable transgenic mice via intracytoplasmic ahESCs injection (ICAI) was very low, which may correlate with the aberrant regulation of imprinted genes. Here we designed to delete the paternal imprinted gene H19 by CRSPR-Cas9 system combined with homologous recombination. The H19 deleted (H19Δ) ahESCs maintained haploidy and genome integrity, expressed pluripotency markers, differentiated into embryoid bodies (EBs), and contributed to chimeras after blastocyst injection. These cells exhibited similar imprinting features with sperm cells, and can produce fertile progenies after ICAI at a high efficiency. More importantly, it is feasible to perform genetic manipulations in H19Δ ahESCs, and the genomic modifications can be properly transmitted to offspring. Our study will benefit the reproductive medicine in curing the hereditary genetic diseases and infertility in the future.
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Project description:Haploid selection within a single ejaculate increases offspring fitness
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Project description:Recent success in the derivation of mouse haploid embryonic stem cells from androgenetic blastocysts (ahESCs) has provided new avenues for the generation of genetically modified animals. However, the efficiency to produce viable transgenic mice via intracytoplasmic ahESCs injection (ICAI) was very low, which may correlate with the aberrant regulation of imprinted genes. Here we designed to delete the paternal imprinted gene H19 by CRSPR-Cas9 system combined with homologous recombination. The H19 deleted (H19Î) ahESCs maintained haploidy and genome integrity, expressed pluripotency markers, differentiated into embryoid bodies (EBs), and contributed to chimeras after blastocyst injection. These cells exhibited similar imprinting features with sperm cells, and can produce fertile progenies after ICAI at a high efficiency. More importantly, it is feasible to perform genetic manipulations in H19Î ahESCs, and the genomic modifications can be properly transmitted to offspring. Our study will benefit the reproductive medicine in curing the hereditary genetic diseases and infertility in the future. The copy number variations of the two H19Î ahESC lines were analyzed by the SurePrint G3 Mouse CGH 4Ã180 K microarrays (Agilent). Wild-type OG-3 ahESCs were used as reference.
2015-11-05 | E-GEOD-67563 | biostudies-arrayexpress
Project description:Doubled haploids (DHs) fix traits from hybrids in one generation. DH induction includes two changes in ploidy levels typically associated with variation in DNA methylation. However, DNA methylation patterns in DH plants and their biological significance are largely unknown. We generated three DH lines in Arabidopsis thaliana by crossing a haploid inducer to the accession Col-0, thus removing tissue culture and hybridization as a variable. DH induction produced thousands of differentially DNA methylated regions (DMRs), most of which were stochastic. Both haploidization and colchicine-induced genome duplication produced DMRs; the former mainly yielded DMRs at non-CG contexts, whereas the latter affected differential gene body methylation. Spontaneous genome doubling of haploid plants also induced DMRs in greater numbers than self-propagation. Our results provide the first evidence that haploid induction and genome doubling result in differential DNA methylation, offering a novel approach to induce epialleles.
2022-12-31 | GSE206313 | GEO
Project description:Mammalian haploid embryonic stem cells (haESCs) provide new possibilities for large-scale genetic screens because they bear only one copy of each chromosome. However, haESCs are prone to spontaneous diploidization through unknown mechanisms. Here, we report that a small molecule combination could restrain mouse haESCs from diploidization by impeding exit from naïve pluripotency and by shortening the S-G2/M phases. Combined with 2i and PD166285, our chemical cocktail could maintain haESCs in the haploid state for at least five weeks without fluorescence-activated cell sorting (FACS) enrichment of haploid cells. Taken together, we established an effective chemical approach for long-term maintenance of haESCs, and highlighted that proper cell cycle progression was critical for the maintenance of haploid state.
2017-10-31 | GSE104519 | GEO
Project description:INTRODUCTION:Kilohertz frequency spinal cord stimulation (kHz-SCS) deposits significantly more power in tissue compared to SCS at conventional frequencies, reflecting increased duty cycle (pulse compression). We hypothesize kHz-SCS increases local tissue temperature by joule heat, which may influence the clinical outcomes. METHODS:To establish the role of tissue heating in KHZ-SCS, a decisive first step is to characterize the range of temperature changes expected during conventional and KHZ-SCS protocols. Fiber optic probes quantified temperature increases around an experimental SCS lead in a bath phantom. These data were used to verify a SCS lead heat-transfer model based on joule heat. Temperature increases were then predicted in a seven-compartment (soft tissue, vertebral bone, fat, intervertebral disc, meninges, spinal cord with nerve roots) geometric human spinal cord model under varied parameterization. RESULTS:The experimentally constrained bio-heat model shows SCS waveform power (waveform RMS) determines tissue heating at the spinal cord and surrounding tissues. For example, we predict temperature increased at dorsal spinal cord of 0.18-1.72?°C during 3.5?mA peak 10?KHz stimulation with a 40-10-40 ?s biphasic pulse pattern, 0.09-0.22?°C during 3.5?mA 1?KHz 100-100-100 ?s stimulation, and less than 0.05?°C during 3.5?mA 50?Hz 200-100-200 ?s stimulation. Notably, peak heating of the spinal cord and other tissues increases superlinearly with stimulation power and so are especially sensitive to incremental changes in SCS pulse amplitude or frequency (with associated pulse compression). Further supporting distinct SCS intervention strategies based on heating; the spatial profile of temperature changes is more uniform compared to electric fields, which suggests less sensitivity to lead position. CONCLUSIONS:Tissue heating may impact short and long-term outcomes of KHZ-SCS, and even as an adjunct mechanism, suggests distinct strategies for lead position and programming optimization.
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Project description:Diploid and haploid strains often exhibit different tolerance to variety of stresses. Transcriptome of acclimation to ethanol stress in diploid and haploid strain of Saccharomyces cerevisiae was analyzed. We analyzed transcriptome profiles of diploid and haploid strains in the presence of ethanol.
2010-01-30 | GSE20108 | GEO