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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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Updated: Aug 17, 2025

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Massively Parallel CRISPR-Based Genetic Perturbation Screening at Single-Cell Resolution.

Junyun Cheng1, Gaole Lin1, Tianhao Wang1

  • 1Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, China.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|December 12, 2022
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Summary

Single-cell CRISPR screening (scCRISPR) offers powerful insights into gene function and biological regulation at an unprecedented resolution. This review details scCRISPR advancements, guiding researchers in selecting optimal methods for their studies.

Keywords:
CRISPR/Casgenetic screeningimaging-based phenotypingmultiplexed perturbationssingle-cell sequencing

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Clustered regularly interspaced short palindromic repeats (CRISPR)-based genetic screening is a key tool for functional genomics.
  • Single-cell CRISPR screening (scCRISPR) integrates CRISPR technology with single-cell analysis for high-resolution biological dissection.
  • scCRISPR enables mass perturbation of cells followed by high-content phenotyping.

Purpose of the Study:

  • To review technical breakthroughs and advancements in various single-cell CRISPR screening methodologies.
  • To analyze the merits and limitations of different scCRISPR approaches.
  • To provide an application-oriented guide for researchers selecting scCRISPR techniques.

Main Methods:

  • scCRISPR methods are categorized by their integration with distinct single-cell technologies: RNA-seq, ATAC-seq, proteome probing, and imaging.
  • Cells are perturbed using pooled CRISPR libraries and annotated with sgRNA-specific barcodes or directly detectable sgRNAs.
  • High-content phenotyping is performed on perturbed single cells.

Main Results:

  • scCRISPR has successfully deciphered complex genotype-phenotype relationships.
  • These techniques have elucidated genetic regulations, tumor biology, and neuropathological mechanisms.
  • The review systematically summarizes advancements, merits, and limitations of diverse scCRISPR methods.

Conclusions:

  • Single-cell CRISPR screening represents a significant technical breakthrough in functional genomics.
  • The diverse methodologies offer powerful tools for dissecting regulatory networks in complex biological systems.
  • An application-oriented guide is provided to assist researchers in choosing appropriate scCRISPR strategies.