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Single Cell RNA Sequencing

Single Cell RNA sequencing is a technique used in the field of Genomics

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patty
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26 views13 pages

Single Cell RNA Sequencing

Single Cell RNA sequencing is a technique used in the field of Genomics

Uploaded by

patty
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Journal of Experimental & Clinical Cancer Research

Genetic and Genome


Engineering

Zhang et al. Journal of Experimental & Clinical Cancer Research (2021) 40:81 https://doi.org/10.1186/s13046-021-01874-1
Abstract
Popular and reliable method.
Examine RNA profile of unique
cells.
Identify differenetial gene
expression and epigenetic factors
caused by mutations.
Reveal heterogeniety of tumor cells.
Monitor tumor development.
Study intercellular communication.
Gain better understanding of
complex tissues.
Abstract

RNA cDNA PCR/IVT Sequencing

(1) Isolation of single cells


(2) Reverse transcription
(3) cDNA amplification
(4) Sequencing library construction
Abstract
Isolation of single cells mainly
includes:
Cell selection,
Dilution of cell suspension
Laser microdissection
(LCM)
Fluorescence-activated cell
sorting (FACS)
Microfluidic/microplate
methodologY.
FACS is the most
commonly used method.
Abstract
Single-cell RNA Sequencing (scRNA-seq) Role of scRNA-seq in
Tumor Research
Analyzes transcriptomes of complex
tissues at single-cell resolution.
Identifies differential gene expression Tumor Future
and epigenetic mutations in unicellular Heterogeneity Implications
genomes.
Discovers new cell-specific markers and Immune System
cell types. Analysis
Endogenous and exogenous environmental factors
Tumor development
Genetic mutations Limitless proliferation
Metastasis
Angiogenesis

Malignant tumor
multi-stage gene
mutations & accumulation
Role of Tumor Tumor microenvironment
Microenvironment
(TME)
(TME)
Tumor Complexity & Immune
Immune Landscape
Cellular Interactions Heterogeneity Mapping
Heterogeneity in Tumor Ecosystems
Cell types in tumors: Malignant, immune, and
stromal cells and their interaction is mediated by
ligand-receptor signaling, autocrine/paracrine Role of scRNA-seq
modes, and other mechanisms. in Tumor
This influence morphology, proliferation, invasion, Metastasis
metastasis, and tumor microenvironment (TME).

Metastatic Clinical
Phenotype Insights
Significance of scRNA-seq
Evaluate transcriptomes at single-cell resolution reveals:
-Rare cell mutations
-Key mutations in tumor progression & heterogeneity
Track tumor heterogeneity dynamics over time
Enhances understanding of:
-Tumor-immune interactions
-Invasive/metastatic abilities and immune cell infiltration
Commercial platforms
10X Genomics
BD Rhapsody (integrated
processes, fast sample
handling).
CITE-seq: Integrates transcriptomics CROP-seq: Links CRISPR screening to
with protein expression profiling. transcriptome analysis for functional studies.

Combines transcriptomics
with other technologies

Rare mutations and


Immunotherapy Applications of heterogeneity
scRNA-seq

Reveals immune cell


infiltration
Role of scRNA-seq in Tumor
Heterogenicity and Evolution
Case Studies
Somatic Mutation
Tumor
Base pair replacements Mapping of gene expression
Insertions, deletions Heterogeneity & differences between primary
Chromosomal and metastatic tumors
Evolution
rearrangements Provide insights into
Copy number variations. metastasis & identifying
potential therapeutic targets.
Tumor Development

Identify gene mutations


Identify expression patterns
Integration of scRNA-seq with Epigenetics

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