• Title/Summary/Keyword: Transcriptomic profiling

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Recent advances in spatially resolved transcriptomics: challenges and opportunities

  • Lee, Jongwon;Yoo, Minsu;Choi, Jungmin
    • BMB Reports
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    • v.55 no.3
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    • pp.113-124
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    • 2022
  • Single-cell RNA sequencing (scRNA-seq) has greatly advanced our understanding of cellular heterogeneity by profiling individual cell transcriptomes. However, cell dissociation from the tissue structure causes a loss of spatial information, which hinders the identification of intercellular communication networks and global transcriptional patterns present in the tissue architecture. To overcome this limitation, novel transcriptomic platforms that preserve spatial information have been actively developed. Significant achievements in imaging technologies have enabled in situ targeted transcriptomic profiling in single cells at single-molecule resolution. In addition, technologies based on mRNA capture followed by sequencing have made possible profiling of the genome-wide transcriptome at the 55-100 ㎛ resolution. Unfortunately, neither imaging-based technology nor capture-based method elucidates a complete picture of the spatial transcriptome in a tissue. Therefore, addressing specific biological questions requires balancing experimental throughput and spatial resolution, mandating the efforts to develop computational algorithms that are pivotal to circumvent technology-specific limitations. In this review, we focus on the current state-of-the-art spatially resolved transcriptomic technologies, describe their applications in a variety of biological domains, and explore recent discoveries demonstrating their enormous potential in biomedical research. We further highlight novel integrative computational methodologies with other data modalities that provide a framework to derive biological insight into heterogeneous and complex tissue organization.

Transcriptomic Profiling of Resistant and Susceptible Soybean Genotypes Following Inoculation of Phytophthora sojae

  • Hee Jin You;Jeongsun Lee;Sungwoo Lee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.188-188
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    • 2022
  • Our previous study identified a resistance locus to Phytophthora sojae (isolate 2457) in an interval of 3.8-4.7 Mbp on chromosome 3 via genetic mapping using a 'Daepung'×'Daewon' recombinant inbred population. Since differential gene expression between Daepung (susceptible) and Daewon (resistant) after inoculation of P. sojae is unknown, RNA sequencing was carried out to compare transcriptomic changes between the two genotypes following inoculation with P. sojae isolate 2457. The two varieties were inoculated using hypocotyl inoculation at the VC stage and stem tissue of 1 cm above and below of the inoculated site were sampled at 0, 6, 12 hours after inoculation (hai), respectively. Differentially expressed genes (DEGs) under same cultivar in different time point and Daepung vs. Daewon in same time point were investigated. In comparison of Daepung vs. Daewon at 12 hai, a total of 3,513 DEGs were identified, including two nucleotide-binding site-leucine rich repeat (NBS-LRR) genes (Glyma.03g034800 and Glyma.03g034900) that are located in the previously reported resistance locus on chromosome 3. In addition, 14,966 DEGs were detected between 0 vs. 6 hai, containing one of candidate genes (Glyma.03g035300). This gene was upregulated by up to 4-fold in Daewon and Daepung. Additional results will be further discussed in the presentation. This study will provide valuable information for soybean crop improvement.

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Whole Transcriptomic Analysis of Bacillus anthracis during Hydrogen Peroxide Decontamination (과산화수소 제독 과정에서의 탄저균 전사체 분석)

  • Kim, Sang Hoon;Kim, Se Kye;Jung, Kyoung Hwa;Yoon, Sung Nyo;Kim, Yun Ki;Kim, Min Cheol;Ryu, Sam Gon;Lee, Hae Wan;Chai, Young Gyu
    • Journal of the Korea Institute of Military Science and Technology
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    • v.18 no.4
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    • pp.478-483
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    • 2015
  • Decontamination of biological agents utilizes hydrogen peroxide($H_2O_2$) for its effectiveness and safeness. Bacillus anthracis is a major target for $H_2O_2$ decontamination. To assess the effect of $H_2O_2$ on B. anthracis and identify biomarkers for decontamination, whole transcriptomic profiling of $H_2O_2$-treated B. anthracis was performed. Here we identified deregulation in stress response genes, transcription factors and cellular homeostasis genes. We also found that expression of antisense RNAs increased in B. anthracis during decontamination. We postulate that B. anthracis prioritizes survival and adaptation in response to $H_2O_2$ treatment by changing its gene expression pattern.

Transcriptomic analysis of the liver in aged laying hens with different intensity of brown eggshell color

  • Han, Gi Ppeum;Kim, Jun-Mo;Kang, Hwan Ku;Kil, Dong Yong
    • Animal Bioscience
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    • v.34 no.5
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    • pp.811-823
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    • 2021
  • Objective: Eggshell color is an important indicator of egg quality for consumers, especially for brown eggs. Various factors related to laying hens and their environment affect brown eggshell coloration. However, there have been no studies investigating hepatic functions of laying hens with variable intensity of brown eggshell color. Therefore, this study was aimed to identify potential factors affecting brown eggshell coloration in aged laying hens at the hepatic transcriptomic level. Methods: Five hundred 92-wk-old Hy-line Brown laying hens were screened to select laying hens with different intensity of brown eggshell color based on eggshell color fans. Based on eggshell color scores, hens with dark brown eggshells (DBE; eggshell color fan score = 14.8) and hens with light brown eggshells (LBE; eggshell color fan score = 9.7) were finally selected for the liver sampling. We performed RNA-seq analysis using the liver samples through the paired-end sequencing libraries. Differentially expressed genes (DEGs) profiling was carried out to identify their biological meaning by bioinformatics. Results: A total of 290 DEGs were identified with 196 being up-regulated and 94 being down-regulated in DBE groups as compared to LBE groups. The Kyoto encyclopedia of genes and genomes (KEGG) analysis revealed that these DEGs belong to several biological pathways including herpes simplex infection (toll-like receptor 3 [TLR3], cyclin-dependent kinase 1, etc.) and influenza A (TLR3, radical S-adenosyl methionine domain containing 2, myxovirus [influenza virus] resistance 1, etc.). Genes related to stress response (ceremide kinase like) and nutrient metabolism (phosphoenolpyruvate carboxy-kinase 1, methylmalonic aciduria [cobalamin deficiency] cblB type, glycine receptor alpha 2, solute carrier family 7 member 11, etc.) were also identified to be differentially expressed. Conclusion: The current results provide new insights regarding hepatic molecular functions related to different intensity of brown eggshell color in aged laying hens. These insights will contribute to future studies aiming to optimize brown eggshell coloration in aged laying hens.

Transcriptomic profiling of the maize (Zea mays L.) to drought stress at the seedling stage

  • Moon, Jun-Cheol;Kim, Hyo Chul;Lee, Byung-Moo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.111-111
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    • 2017
  • The development and productivity of maize (Zea mays L.) is frequently impacted by water scarcity, and consequently to increased drought tolerance in a priority target in maize breeding programs. To elucidate the molecular mechanisms of resistance to drought stress in maize, RNA-seq of the public database was used for transcriptome profiling of the seedling stage exposed to drought stress of three levels, such as moderate, severe drought stress and re-watering. In silico analysis of differentially expressed genes (DEGs), 176 up-regulated and 166 down-regulated DEGs was detected at moderated stress in tolerance type. These DEGs was increasing degradation of amino acid metabolism in biological pathways. Six modules based on a total of 4,771 DEGs responses to drought stress by the analysis of co-expression network between tolerance and susceptible type was constructed and showed to similar module types. These modules were discriminated yellow, greenyellow, turquoise, royalblue, brown4 and plum1 with 318, 2433, 375, 183, 1405 and 56 DEGs, respectively. This study was selected 30 DEGs to predicted drought stress response gene and was evaluated expression levels using drought stress treated sample and re-watering sample by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). 23 genes was shown increasing with drought stress and decreasing with re-watering. This study contribute to a better understanding of the molecular mechanisms of maize seedling stage responses to drought stress and could be useful for developing maize cultivar resistant to drought stress.

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Molecular Cloning, Transcriptome Profiling, and Characterization of Histone Genes in the Dinoflagellate Alexandrium pacificum

  • Riaz, Sadaf;Sui, Zhenghong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1185-1198
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    • 2018
  • The nucleosomal organization of chromatin using histone proteins is a fundamental and ubiquitous feature of eukaryotic nuclei, with the major exception of dinoflagellates. Although a number of recent genomic and transcriptomic analyses have detected numerous histone genes in dinoflagellates, little is known about their expression. Here in, we aimed to investigate the expression pattern of histone genes under nutritional stress, and an attempt was made to detect histone expression at the protein level in Alexandrium pacificum. The presence of histones at the mRNA level was confirmed in this study by the amplification, cloning, and sequencing of 10 different genes. Relative expression profiling of these genes under different growth conditions was determined with real-time PCR and revealed considerable levels of histone transcription in nutritionally stressed cells. We were unable to detect the expression of histones at the protein level even after immunodetection and analysis using mass spectrometry, although a histone-like protein was detected as a major nuclear component. A. pacificum expresses multiple variants of histone, and protein sequences revealed both conservation and divergence with respect to other eukaryotes. We concluded that A. pacificum maintained an active transcription of histone genes within the cell, and enhanced expression of histone genes in nutritional stress strongly suggest that histones have functional significance in dinoflagellates, although expression at the protein level was below our current detection limits, which suggests a limited role of histones in DNA packaging. Finally, the plausible regulation of histone expression at the gene and protein levels in A. pacificum is discussed.

De novo transcriptome sequencing and gene expression profiling with/without B-chromosome plants of Lilium amabile

  • Park, Doori;Kim, Jong-Hwa;Kim, Nam-Soo
    • Genomics & Informatics
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    • v.17 no.3
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    • pp.27.1-27.9
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    • 2019
  • Supernumerary B chromosomes were found in Lilium amabile (2n = 2x = 24), an endemic Korean lily that grows in the wild throughout the Korean Peninsula. The extra B chromosomes do not affect the host-plant morphology; therefore, whole transcriptome analysis was performed in 0B and 1B plants to identify differentially expressed genes. A total of 154,810 transcripts were obtained from over 10 Gbp data by de novo assembly. By mapping the raw reads to the de novo transcripts, we identified 7,852 differentially expressed genes (log2FC > |10|), in which 4,059 and 3,794 were up-and down-regulated, respectively, in 1B plants compared to 0B plants. Functional enrichment analysis revealed that various differentially expressed genes were involved in cellular processes including the cell cycle, chromosome breakage and repair, and microtubule formation; all of which may be related to the occurrence and maintenance of B chromosomes. Our data provide insight into transcriptomic changes and evolution of plant B chromosomes and deliver an informative database for future study of B chromosome transcriptomes in the Korean lily.

Transcriptome Analysis and Expression Profiling of Molecular Responses to Cd Toxicity in Morchella spongiola

  • Xu, Hongyan;Xie, Zhanling;Jiang, Hongchen;Guo, Jing;Meng, Qing;Zhao, Yuan;Wang, Xiaofang
    • Mycobiology
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    • v.49 no.4
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    • pp.421-433
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    • 2021
  • Morchella is a genus of fungi with the ability to concentrate Cd both in the fruit-body and mycelium. However, the molecular mechanisms conferring resistance to Cd stress in Morchella are unknown. Here, RNA-based transcriptomic sequencing was used to identify the genes and pathways involved in Cd tolerance in Morchella spongiola. 7444 differentially expressed genes (DEGs) were identified by cultivating M. spongiola in media containing 0.15, 0.90, or 1.50 mg/L Cd2+. The DEGs were divided into six sub-clusters based on their global expression profiles. GO enrichment analysis indicated that numerous DEGs were associated with catalytic activity, cell cycle control, and the ribosome. KEGG enrichment analysis showed that the main pathways under Cd stress were MAPK signaling, oxidative phosphorylation, pyruvate metabolism, and propanoate metabolism. In addition, several DEGs encoding ion transporters, enzymatic/non-enzymatic antioxidants, and transcription factors were identified. Based on these results, a preliminary gene regulatory network was firstly proposed to illustrate the molecular mechanisms of Cd detoxification in M. spongiola. These results provide valuable insights into the Cd tolerance mechanism of M. spongiola and constitute a robust foundation for further studies on detoxification mechanisms in macrofungi that could potentially lead to the development of new and improved fungal bioremediation strategies.