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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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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.
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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Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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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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Updated: Nov 1, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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CRISPR/Cas Technologies Applied to Pseudogenes.

Marianna Vitiello1,2, Laura Poliseno3,4

  • 1Oncogenomics Unit, Core Research Laboratory, ISPRO, Pisa, Italy. mvitiello@ifc.cnr.it.

Methods in Molecular Biology (Clifton, N.J.)
|June 24, 2021
PubMed
Summary
This summary is machine-generated.

Pseudogenes, once dismissed as junk DNA, are now recognized for their biological roles. CRISPR/Cas technology offers novel methods to precisely study and manipulate these pseudogenes and their functions.

Keywords:
CRISPR/Cas systemCRISPRaCRISPRiCas13Cas9Genome editingPseudogeneRCas9RNA ImmunoprecipitationRNA editingRNA pull-downTranscript tracking

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Area of Science:

  • Molecular Biology
  • Genetics
  • Genomics

Background:

  • Pseudogenes were historically considered nonfunctional DNA, often termed "junk DNA" or "transcriptional noise."
  • Emerging evidence highlights pseudogene involvement in critical biological processes, necessitating their detailed study.
  • Pseudogene research is challenging due to high homology with parental genes and their function as noncoding RNAs.

Purpose of the Study:

  • To explore the application of CRISPR/Cas technology for pseudogene research.
  • To demonstrate how CRISPR/Cas can overcome challenges in pseudogene manipulation and study.
  • To present CRISPR/Cas as a versatile tool for pseudogene DNA and RNA editing and analysis.

Main Methods:

  • Genome editing using CRISPR/Cas for pseudogene knock-out (complete elimination) and knock-in (sequence insertion).
  • CRISPR/Cas mediated manipulation of pseudogene transcription (both activation and repression).
  • Targeting pseudogene RNA for expression regulation and using CRISPR/Cas as an RNA Binding Protein system for molecular biology techniques.

Main Results:

  • CRISPR/Cas enables precise genome editing of pseudogenes, including complete removal or specific sequence alterations.
  • The technology allows for controlled manipulation of pseudogene transcription levels.
  • CRISPR/Cas facilitates targeted modification of pseudogene DNA and RNA sequences.
  • CRISPR/Cas functions effectively as an RNA Binding Protein system for various molecular biology applications.

Conclusions:

  • CRISPR/Cas technology provides powerful and versatile solutions for overcoming pseudogene research challenges.
  • This technology facilitates precise manipulation and functional studies of pseudogenes at both DNA and RNA levels.
  • CRISPR/Cas expands the toolkit for pseudogene research, including applications in transcript tracking and live imaging.