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High throughput error correction using dual nucleotide dimer blocks allows direct single-cell nanopore transcriptome sequencing

ABSTRACT: High throughput error correction using dual nucleotide dimer blocks allows direct single-cell nanopore transcriptome sequencing

PROVIDER: PRJNA680369 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13120810_1.fastq.gz	Fastqsanger.gz
	SRR13397411_1.fastq.gz	Fastqsanger.gz
	SRR13397412_1.fastq.gz	Fastqsanger.gz
	SRR13397413_1.fastq.gz	Fastqsanger.gz
	SRR13397414_1.fastq.gz	Fastqsanger.gz

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High throughput error correction using dual nucleotide dimer blocks allows direct single-cell nanopore transcriptome sequencing

Project description:Droplet-based single-cell sequencing techniques have provided unprecedented insight into cellular heterogeneities within tissues. However, these approaches only allow for the measurement of the distal parts of a transcript following short-read sequencing. Therefore, splicing and sequence diversity information is lost for the majority of the transcript. The application of long-read Nanopore sequencing to droplet-based methods is challenging because of the low base-calling accuracy currently associated with Nanopore sequencing. Although several approaches that use additional short-read sequencing to error-correct the barcode and UMI sequences have been developed, these techniques are limited by the requirement to sequence a library using both short- and long-read sequencing. Here we introduce a novel approach termed single-cell Barcode UMI Correction sequencing (scBUC-seq) to efficiently error-correct barcode and UMI oligonucleotide sequences synthesized by using blocks of dimeric nucleotides. The method can be applied to correct both short-read and long-read sequencing, thereby allowing users to recover more reads per cell that permits direct single-cell Nanopore sequencing for the first time. We illustrate our method by using species-mixing experiments to evaluate barcode assignment accuracy and multiple myeloma cell lines to evaluate differential isoform usage and Ewing’s sarcoma cells to demonstrate Ig fusion transcript analysis.

2021-01-31 | GSE162053 | GEO

A cell type specific error correction signal in posterior parietal cortex

Project description:Neurons in posterior parietal cortex contribute to the execution of goal-directed navigation and other decision-making tasks. Although molecular studies have catalogued over fifty cortical cell types, it remains unknown what distinct functions they serve during goal-directed navigation. Here, we identified a molecularly defined subset of somatostatin (Sst) inhibitory neurons that, in mouse posterior parietal cortex, carry a novel cell type-specific error correction signal for navigation. We obtained repeatable experimental access to these cells using an adeno-associated virus (AAV) in which gene expression is driven by an enhancer that functions specifically in a subset of Sst cells. We found that during goal-directed navigation in a virtual environment, this subset of Sst neurons activates in a synchronous pattern that is distinct from the activity of surrounding neurons, including other Sst neurons. Using in vivo two-photon photostimulation and ex vivo paired patch clamp recordings, we show that nearby cells of this Sst subtype excite each other through gap junctions, revealing a self-excitation circuit motif that contributes to the synchronous activity of this cell type. Remarkably, these cells selectively activate as mice execute course corrections for deviations in their virtual heading during navigation toward a reward location, both for self- and experimentally-induced deviations. We propose that this subtype of Sst neurons provides a self-reinforcing and cell type-specific error-correction signal in posterior parietal cortex that may aid the execution and learning of accurate goal-directed navigation trajectories.

2023-05-11 | GSE232200 | GEO

Quantifying sequencing error and effective sequencing depth of liquid biopsy NGS with unique molecular identifier error correction

Project description:Quantifying sequencing error and effective sequencing depth of liquid biopsy NGS with unique molecular identifier error correction

| PRJEB39899 | ENA

A cell type specific error correction signal in posterior parietal cortex

Project description:A cell type specific error correction signal in posterior parietal cortex

| PRJNA971206 | ENA

Proximity-based activation of AURORA A by MPS1 potentiates error correction

Project description:Faithfull cell division relies on mitotic chromosomes becoming bioriented with each pair of sister kinetochores bound to microtubules from opposing spindle poles. Erroneous kinetochore- microtubule attachments often form during early mitosis, but are destabilized through the phosphorylation of outer kinetochore proteins by centromeric AURORA B kinase (ABK) and centrosomal AURORA A kinase (AAK), thus allowing for re-establishment of attachments until biorientation is achieved. MPS1-mediated phosphorylation of NDC80 has also been shown to directly weaken the kinetochore-microtubule interface in yeast. In human cells, MPS1 has been proposed to transiently accumulate at end-on attached kinetochores and phosphorylate SKA3 to promote microtubule release. Whether MPS1 directly targets NDC80 and/or promotes the activity of AURORA kinases in metazoans remains unclear. Here, we report a novel mechanism involving communication between kinetochores and centrosomes, wherein MPS1 acts upstream of AAK to promote error correction. MPS1 on pole-proximal kinetochores phosphorylates the C-lobe of AAK thereby increasing its activation at centrosomes. This proximity-based activation ensures the establishment of a robust AAK activity gradient that locally destabilizes mal-oriented kinetochores near spindle poles. Accordingly, MPS1 depletion from Drosophila cells causes severe chromosome misalignment and erroneous kinetochore-microtubule attachments, which can be rescued by tethering either MPS1 or constitutively active AAK mutants to centrosomes. Proximity-based activation of AAK by MPS1 also occurs in human cells to promote AAK-mediated phosphorylation of the NDC80 N-terminal tail. These findings uncover an MPS1-AAK cross-talk that is required for efficient error correction, showcasing the ability of kinetochores to modulate centrosome outputs to ensure proper chromosome segregation.

2025-04-03 | PXD056173 | Pride

Error-correction enables use of Oxford Nanopore technology for reference-free transcriptome analysis

Project description:Error-correction enables use of Oxford Nanopore technology for reference-free transcriptome analysis

| PRJEB34849 | ENA

DNA replication-mediated error correction of ectopic CENP-A deposition maintains centromere identity

Project description:DNA replication-mediated error correction of ectopic CENP-A deposition maintains centromere identity

| PRJNA436748 | ENA

DNA replication-mediated error correction of ectopic CENP-A deposition maintains centromere identity

Project description:Chromatin assembled with histone H3 variant CENP-A is the heritable epigenetic determinant of human centromere identity. Using genome-wide mapping and reference models for 23 human centromeres, CENP-A is shown in early G1 to be assembled into nucleosomes within megabase, repetitive a-satellite DNAs at each centromere and onto 11,390 sites on the chromosome arms. Centromere-bound CENP-A is found to be quantitatively maintained during DNA replication by coordinated action of the MCM2 helicase, CAF1, HJURP, and the CCAN network of constitutive centromere components. CCAN serves to tether CENP-A removed by MCM2, thereby enabling local reassembly onto both daughter centromeres with identical DNA sequence preferences and nucleosome phasing as the loading in G1 and independent of CENP-B. Conversely, without CCAN-mediated tethering, DNA replication removes CENP-A from sites on the chromosome arms. Our data identify an MCM2/CAF1/HJURP- and CCAN-dependent error correction mechanism that acts in S-phase to maintain CENP-A-dependent centromere identity.

2019-03-20 | GSE111381 | GEO

Made in error

Project description:Made in error

| PRJEB31468 | ENA

Project description:Error correction tool benchmark

| PRJNA504496 | ENA