Shortening the half-life of Cas9 maintains its gene editing ability and reduces neuronal toxicity
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ABSTRACT:
PROVIDER: PRJNA503401 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:Virus-mediated expression of CRISPR/Cas9 has been actively used for genome editing in animal models of genetic disorders, including neurological diseases, but the consequences of overexpressing bacterial Cas9 in the mammalian brain remain unknown. Through RNA-seq analysis, we found that virus-mediated expression of Cas9 caused systematic changes in genes involved in neuronal functions. We also generated a short-lived version of Cas9, which maintains its genome editing capacity, but significantly alleviates neurotoxicity caused by overexpressed Cas9. Thus, modification of Cas9 by shortening its half-life would help develop CRISPR/Cas9-based therapeutic approaches for treating genetic neurological disorders.
2018-12-04 | GSE122072 | GEO
Project description:Virus-mediated expression of CRISPR/Cas9 is commonly used for genome editing in animal brains to model or treat neurological diseases, but the potential neurotoxicity of overexpressing bacterial Cas9 in the mammalian brain remains unknown. Through RNA sequencing (RNA-seq) analysis, we find that virus-mediated expression of Cas9 influences the expression of genes involved in neuronal functions. Reducing the half-life of Cas9 by tagging with geminin, whose expression is regulated by the cell cycle, maintains the genome editing capacity of Cas9 but significantly alleviates neurotoxicity. Thus, modification of Cas9 by shortening its half-life can help develop CRISPR/Cas9-based therapeutic approaches for treating neurological disorders.
| S-EPMC6314484 | biostudies-literature
Project description:To help elucidate mechanistic differences between lengthening and shortening muscle contractions we compared changes in gene expression within 24 h of an acute bout of each type of contraction in the quadriceps. Three healthy young men performed shortening contractions with one leg while the contralateral leg performed lengthening contractions. Biopsies were taken from both legs before exercise, 3, 6, and 24 h afterwards. Expression profiling was performed using a custom-made Affymetrix MuscleChip containing probesets of approximately 3,300 known genes and ESTs expressed in skeletal muscle. We identified 49 genes differentially regulated between the lengthening and shortening contractions. Using unsupervised hierarchical clustering, we identified four distinct clusters, three of which corresponded to unique functional categories (protein synthesis, stress response/early growth, and sarcolemmal structure). The results suggest distinct molecular pathways as early as 3 h post exercise. The molecular differences might contribute to mechanisms underlying the physiological adaptations seen with training using the two modes of exercise. Keywords: Time Series Design The hypothesis of the current study was that genes involved in hypertrophy will be differentially regulated between the two types of contractions.
2007-03-27 | E-GEOD-7286 | biostudies-arrayexpress
Project description:To help elucidate mechanistic differences between lengthening and shortening muscle contractions we compared changes in gene expression within 24 h of an acute bout of each type of contraction in the quadriceps. Three healthy young men performed shortening contractions with one leg while the contralateral leg performed lengthening contractions. Biopsies were taken from both legs before exercise, 3, 6, and 24 h afterwards. Expression profiling was performed using a custom-made Affymetrix MuscleChip containing probesets of approximately 3,300 known genes and ESTs expressed in skeletal muscle. We identified 49 genes differentially regulated between the lengthening and shortening contractions. Using unsupervised hierarchical clustering, we identified four distinct clusters, three of which corresponded to unique functional categories (protein synthesis, stress response/early growth, and sarcolemmal structure). The results suggest distinct molecular pathways as early as 3 h post exercise. The molecular differences might contribute to mechanisms underlying the physiological adaptations seen with training using the two modes of exercise. Keywords: Time Series Design
2007-03-27 | GSE7286 | GEO
Project description:We have identified NANOS2 bound targets transcriptome wide by employing CRAC and RNA-seq in mouse SSCs. We have determined the average mRNA half-life in SSCs by SLAM-seq. By comparing CRAC and SLAM-seq datasets, we have demonstrated that NANOS2 binding reduces mRNA stability in SSCs.
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Project description:Despite decades of research, predicting where a transcription factor (TF) binds using its sequences alone continues to be an outstanding problem. Here, our thermodynamic model of Gal4p-DNA binding revealed a new mechanism of binding site selection in which Gal4p can bind to promoters that lacks it consensus site. Further investigation revealed that these sequences that Gal4p unexpectedly binds to have a high density of its half site, CGG. Analyses showed the ability of CGG half sites to predict zinc cluster TF in vivo binding and exposed a clear relationship between CGG half sites and zinc cluster binding in multiple binding datasets. To dissect the molecular logic of this half site mode of binding we designed a Sort-seq library containing 6,798 yeast promoter sequences and measured their expression. We learned that there is a linear relationship between CGG half site density and zinc cluster driven expression in vivo. Additionally, we found that promoters encoding 10 half sites have the same transcriptional output as a single consensus site and found no clear effects of half site orientation and spacing on strength of activation. Our experiments demonstrate that this novel mode of half site binding is widespread across the yeast genome and expands our knowledge about TF binding specificity that can be applied to higher eukaryotes.
2022-11-04 | GSE216839 | GEO
Project description:Despite decades of research, predicting where a transcription factor (TF) binds using its sequences alone continues to be an outstanding problem. Here, our thermodynamic model of Gal4p-DNA binding revealed a new mechanism of binding site selection in which Gal4p can bind to promoters that lacks it consensus site. Further investigation revealed that these sequences that Gal4p unexpectedly binds to have a high density of its half site, CGG. Analyses showed the ability of CGG half sites to predict zinc cluster TF in vivo binding and exposed a clear relationship between CGG half sites and zinc cluster binding in multiple binding datasets. To dissect the molecular logic of this half site mode of binding we designed a Sort-seq library containing 6,798 yeast promoter sequences and measured their expression. We learned that there is a linear relationship between CGG half site density and zinc cluster driven expression in vivo. Additionally, we found that promoters encoding 10 half sites have the same transcriptional output as a single consensus site and found no clear effects of half site orientation and spacing on strength of activation. Our experiments demonstrate that this novel mode of half site binding is widespread across the yeast genome and expands our knowledge about TF binding specificity that can be applied to higher eukaryotes.
2022-11-04 | GSE216838 | GEO
Project description:Global RNA half lives in cyanobacteria
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Project description:Gene expression changes of UC-model organoids derive from human colon indcued by telomere enlongation was assessed by Madecassoside treatemt (telomerase activator). Gene expression changes of human colon organoids induced by telomere shortening was assessd by MST-312 treatment (telomerase inhibitor).
2021-06-24 | GSE175994 | GEO
Project description:Sensing of environmental cues is crucial for cell survival. To adapt to changes in their surroundings cells need to tightly control the repertoire of genes expressed at any time. Regulation of translation is key, especially in organisms in which transcription is hardly controlled, like Trypanosoma brucei. In this study we describe the shortening of the bulk of the cellular tRNAs during stress at the expense of the conserved 3’ CCA-tail. This tRNA shortening is specific for nutritional stress and renders tRNAs unsuitable substrates for translation. We uncovered the nuclease LCCR4 (Tb927.4.2430), a homologue of the conserved deadenylase Ccr4, as being responsible for tRNA trimming. Once optimal growth conditions are restored tRNAs are rapidly repaired by the trypanosome tRNA nucleotidyltransferase thus rendering the recycled tRNAs amenable for translation. This mechanism represents a fast and efficient way to repress translation during stress, allowing quick reactivation with a low energy input.
2021-01-13 | PXD021540 | Pride