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Related Concept Videos

RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues
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Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues

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A targeted sequencing extension for transcript genotyping in single-cell transcriptomics.

Lies Van Horebeek1, Margaux David1, Nina Dedoncker1

  • 1Laboratory for Neuroimmunology, Department of Neurosciences, Leuven Brain Institute, KU Leuven, Leuven, Belgium.

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|September 11, 2023
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Summary
This summary is machine-generated.

This study introduces a targeted sequencing method to combine genotyping with single-cell RNA sequencing. This approach enables the study of genetic variants

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Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Single-cell RNA sequencing (scRNA-seq) lacks methods for high-throughput genotyping of specific genomic loci.
  • Understanding the functional impact of genetic variants at the single-cell level is crucial.

Purpose of the Study:

  • To develop and evaluate a targeted sequencing extension for droplet-based scRNA-seq.
  • To integrate transcript-level genotype information with gene expression data.
  • To enable the study of genetic variant impact on cellular transcriptomes.

Main Methods:

  • Extension of droplet-based scRNA-seq with targeted sequencing.
  • Processing of scRNA-seq and targeted sequencing data.
  • Evaluation of technical parameters (cDNA input, amplification cycles, sequencing depth).

Main Results:

  • Demonstrated a straightforward targeted sequencing extension for scRNA-seq.
  • Assessed the influence of technical parameters on transcript detection.
  • Showcased applications in somatic variant analysis and allele-specific expression studies.

Conclusions:

  • The targeted sequencing extension facilitates simultaneous analysis of genetic variants and transcriptomes.
  • This method aids in investigating the functional effects of genetic variation in single cells.
  • Potential to advance understanding of genotype-phenotype relationships at single-cell resolution.