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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/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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DNA-only Transposons02:57

DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
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Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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Related Experiment Video

Updated: Nov 16, 2025

Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology
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Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology

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Transposon-Associated CRISPR-Cas System: A Powerful DNA Insertion Tool.

Wang Ma1, Ying-Shuang Xu1, Xiao-Man Sun1

  • 1School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, 2 Xuelin Road, Qixia District, Nanjing, People's Republic of China.

Trends in Microbiology
|February 22, 2021
PubMed
Summary

The transposon-associated CRISPR-Cas system enables precise genetic modifications like knockins and knockouts. This advanced genome editing tool works independently of host repair, expanding its utility across diverse cell types.

Keywords:
DNA insertionmicrobial community genome editingtargeted genome editingtransposable elementstransposon-associated CRISPR-Cas system

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas systems are revolutionary tools for genome engineering.
  • Existing methods often rely on host-cell DNA repair pathways, limiting their efficiency and applicability.
  • Transposon-associated systems offer an alternative mechanism for genetic manipulation.

Purpose of the Study:

  • To evaluate the efficacy of the transposon-associated CRISPR-Cas system for targeted genetic manipulation.
  • To demonstrate the system's ability to perform genomic knockins and knockouts.
  • To highlight the system's independence from host-cell repair machinery.

Main Methods:

  • Utilizing a transposon-mediated delivery system for CRISPR-Cas components.
  • Implementing targeted guide RNAs for specific genomic loci.
  • Assessing editing outcomes through molecular analysis.

Main Results:

  • Successful targeted genomic knockins and knockouts were achieved using the transposon-associated CRISPR-Cas system.
  • Editing efficiency was demonstrated across various cell types.
  • The system's functionality was confirmed to be independent of endogenous DNA repair pathways.

Conclusions:

  • The transposon-associated CRISPR-Cas system represents a significant advancement in genome editing technology.
  • Its ability to function independently of host repair broadens the scope of genome editing applications.
  • This system offers a versatile and efficient tool for genetic manipulation in diverse cellular contexts.