Cell subset annotation method based on single cell transcriptome sequencing

A technology of transcriptome sequencing and cell subgroups, which is applied in the field of bioinformatics analysis, can solve problems such as annotation of single cell subgroups, and achieve the effect of comprehensive cell subgroups and avoiding subjective consciousness

Active Publication Date: 2021-04-23
广州华银医学检验中心有限公司 +1
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  • Abstract
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  • Application Information

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Problems solved by technology

[0007] In order to solve the problems existing in the prior art, the present invention provides a cell subgroup annotation method based on single-cell transcriptome sequencing, which is mainly used for annotation of standard single-cell transcriptome cell subgroups of 10x Genomics platform data, and solves the problem of single-cell transcriptome cell subgroup annotation. The problem of annotation of cell subgroups enables single-cell sequencing data to support cell annotation based on gene expression profiles and / or cell marker genes after routine analysis, realizing the organic combination of different annotation methods, and obtaining the distribution of cell types and correlation information

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  • Cell subset annotation method based on single cell transcriptome sequencing
  • Cell subset annotation method based on single cell transcriptome sequencing
  • Cell subset annotation method based on single cell transcriptome sequencing

Examples

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Embodiment 1

[0037] Example 1 Cell Segmentation and Annotation of Normal Chorionic Cells

[0038] A cell subpopulation annotation method based on single-cell transcriptome sequencing, including the following steps:

[0039] S1. 10x barcode UMI identification: The off-machine sequencing data of the 10x genomics platform database is a fastq sequence, and the fastq sequence with the same ID number includes 3 parts: barcode+UMI+mRNA sequence, use the software cellrangercount to distinguish the sequence through the barcode sequence Source cells, quantify gene expression through UMI sequence, and use 3' end mRNA sequence for gene identification;

[0040] S2. Genome comparison: use the STAR algorithm to compare the fastq sequence obtained by sequencing to the reference genome, and locate the measured sequence to the corresponding gene;

[0041] S3. Gene expression profile construction: including the following steps: 1) Data integration and data volume normalization: when samples from multiple li...

Embodiment 2

[0052] Example 2 Cell Segmentation and Annotation of Diseased Chorionic Cells

[0053] A cell subpopulation annotation method based on single-cell transcriptome sequencing, including the following steps:

[0054] S1. 10x barcode UMI identification: The off-machine sequencing data of the 10x genomics platform database is a fastq sequence, and the fastq sequence with the same ID number includes 3 parts: barcode+UMI+mRNA sequence, use the software cellrangercount to distinguish the sequence through the barcode sequence Source cells, gene expression quantification by UMI sequence, gene identification by 3' end mRNA sequence;

[0055] S2. Genome comparison: use the STAR algorithm to compare the fastq sequence obtained by sequencing to the reference genome, and locate the measured sequence to the corresponding gene;

[0056] S3. Gene expression profile construction: including the following steps: 1) Data integration and data volume normalization: when samples from multiple librarie...

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Abstract

The invention provides a cell subset annotation method based on single cell transcriptome sequencing. The cell subset annotation method comprises the following steps: 1) 10x barcode UMI identification; 2) genome comparison; 3) gene expression profile comparison; 4) low-quality cell filtering and data homogenization; 5) cell population clustering; 6) marker gene extraction; 7) cell subset annotation. The invention belongs to the technical field of biological information analysis, and provides a cell subset annotation method based on single-cell transcriptome sequencing, which solves the problem of single-cell subset annotation, so that single-cell sequencing data can support cell annotation according to a gene expression profile and / or a cell Marker gene after conventional analysis. Therefore, organic combination of different annotation methods is realized, and the distribution condition and related information of cell types are obtained.

Description

technical field [0001] The invention belongs to the technical field of biological information analysis, and in particular relates to a cell subgroup annotation method based on single-cell transcriptome sequencing. Background technique [0002] With the rapid development of sequencing technology, especially high-throughput sequencing technology, people have a deeper understanding of the relationship between genomic variation / gene expression differences and phenotypes. However, the traditional Bulk RNA sequencing method is to sequence the cell collection, and the specific information of a single cell is often covered up, resulting in the loss of many important information. [0003] Single-cell transcriptome sequencing technology is applied to genome-wide gene regulatory networks in individual cells, especially in highly heterogeneous cell populations, such as stem cells, early embryonic cells, and tumor cells. The clinical application of this technology can continuously track...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G16B30/10G16B40/00G16B50/00
CPCG16B30/10G16B40/00G16B50/00
Inventor 范文涛王勇斯张盼玉温韵洁何丹
Owner 广州华银医学检验中心有限公司
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