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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

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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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CRISPR and crRNAs02:53

CRISPR and crRNAs

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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RNA Editing02:23

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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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Comparing Mitochondrial, Chloroplast, and Prokaryotic Genomes02:16

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The present-day mitochondrial and chloroplast genomes have retained some of the characteristics of their ancestral prokaryotes and also have acquired new attributes during their evolution within eukaryotic cells. Like prokaryotic genomes, mitochondrial and chloroplast genomes neither bind with histone-like proteins nor show complex packaging into chromosome-like structures, as observed in eukaryotes. Unlike mitotic cell divisions observed in eukaryotic cells, mitochondria and chloroplasts...
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Prokaryotic Transcriptional Activators and Repressors01:58

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The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
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Related Experiment Video

Updated: Jan 31, 2026

Microinjection of Corn Planthopper, Peregrinus maidis, Embryos for CRISPR/Cas9 Genome Editing
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A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants.

Xu Tang1, Zhaohui Zhong1, Qiurong Ren1

  • 1Department of Biotechnology, School of Life Science and Technology, Center for Informational Biology, University of Electronic Science and Technology of China, Chengdu, China.

Methods in Molecular Biology (Clifton, N.J.)
|January 6, 2019
PubMed
Summary

This study introduces a novel single transcript unit CRISPR-Cas9 system for multiplex genome editing. This approach simplifies vector construction by using a single Pol II promoter for multiple guide RNAs, overcoming limitations of conventional methods.

Keywords:
CRISPR-Cas9Multiplex genome editingSTU

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The CRISPR-Cas9 system is a powerful tool for genome editing, enabling targeted modifications.
  • Multiplex genome editing, or simultaneous editing of multiple genomic sites, is crucial for complex genetic studies.
  • Conventional methods for multiplex CRISPR-Cas9 often rely on expressing multiple single-guide RNAs (sgRNAs) using Pol III promoters, which complicates vector construction.

Purpose of the Study:

  • To develop a simplified and efficient system for multiplex genome editing.
  • To overcome the challenges associated with vector construction in conventional multiplex CRISPR-Cas9 systems.
  • To present a novel approach utilizing a single transcript unit (STU) CRISPR-Cas9 expression system.

Main Methods:

  • Development of a single transcript unit (STU) CRISPR-Cas9 expression system.
  • Utilizing a single Pol II promoter to drive the expression of multiple sgRNAs.
  • Demonstration of multiplex genome editing capabilities with the novel system.

Main Results:

  • The STU CRISPR-Cas9 system enables multiplex genome editing.
  • This novel system is driven by a single Pol II promoter, simplifying expression.
  • The approach facilitates easier vector construction compared to conventional methods.

Conclusions:

  • The developed STU CRISPR-Cas9 system offers a novel and efficient strategy for multiplex genome editing.
  • This method simplifies the process by using a single expression unit and promoter.
  • The findings present a significant advancement for applications requiring simultaneous modification of multiple genomic loci.