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Related Concept Videos

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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CRISPR and crRNAs02:53

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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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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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The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
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Eukaryotic RNA Polymerases00:58

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RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
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RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
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Related Experiment Video

Updated: Jan 27, 2026

Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9
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RAPID-DASH: fast and efficient assembly of guide RNA arrays for multiplexed CRISPR-Cas9 applications.

Asfar Lathif Salaudeen1, Nicholas Mateyko1, Carl G de Boer2

  • 1Genome Science and Technology Graduate Program, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada.

Synthetic Biology (Oxford, England)
|January 26, 2026
PubMed
Summary

Researchers developed a fast CRISPR-Cas9 method to create guide RNA (gRNA) arrays for multiplexed gene editing. This technique enables efficient simultaneous targeting of multiple genomic sites, accelerating genetic research and combinatorial perturbation studies.

Keywords:
Golden Gate assemblygRNA arraygenome engineeringmultiplexingpolymerase cycling assemblyCRISPR-Cas9

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas9 technology allows precise genome editing.
  • Simultaneous targeting of multiple genomic loci requires complex guide RNA (gRNA) designs.
  • Current methods for constructing gRNA arrays can be time-consuming and inefficient.

Purpose of the Study:

  • To develop a streamlined and efficient method for rapidly constructing guide RNA (gRNA) arrays.
  • To enable the simultaneous targeting of up to 10 genomic loci using CRISPR-Cas9.
  • To facilitate combinatorial perturbation research through economical and rapid gRNA array assembly.

Main Methods:

  • A novel method for assembling gRNA arrays with up to 10 gRNA units in a single day.
  • Demonstration of functional activity of gRNA arrays across all positions.
  • Incorporation of gRNA libraries to combine scalability and multiplexing.

Main Results:

  • Successful rapid construction of gRNA arrays within one day.
  • Consistent and robust functional activity of all incorporated gRNAs, regardless of position.
  • Demonstrated scalability and multiplexing capabilities for combinatorial studies.

Conclusions:

  • The developed method provides a significant advancement for constructing multiplexed CRISPR-Cas9 systems.
  • This approach accelerates the pace of combinatorial perturbation research by simplifying gRNA array assembly.
  • A user-friendly web tool is provided to aid in the design of necessary oligonucleotide sequences, promoting wider adoption.