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CRISPR01:59

CRISPR

57.2K
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...
1.4K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.5K
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...
6.5K
CRISPR and crRNAs02:53

CRISPR and crRNAs

18.5K
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...
18.5K
Homologous Recombination02:31

Homologous Recombination

61.9K
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...
61.9K
The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

522
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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Related Experiment Video

Updated: Dec 20, 2025

Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology
10:07

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

Published on: March 16, 2022

2.5K

CRISPR Lights up In Situ Protein Evolution.

Evan M Kerek1, Christopher R Cromwell1, Basil P Hubbard1

  • 1Department of Pharmacology, University of Alberta, Edmonton, AB T6G 2R7, Canada.

Cell Chemical Biology
|May 23, 2020
PubMed
Summary
This summary is machine-generated.

Researchers developed a CRISPR/Cas9 method for directed evolution of mammalian proteins. This approach identified mCRISPRed, a stable fluorescent protein optimized for low pH environments like lysosomes.

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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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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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • Directed evolution is crucial for protein engineering.
  • CRISPR/Cas9 technology offers precise genome editing capabilities.
  • Mammalian protein engineering often requires in situ functional selection.

Purpose of the Study:

  • To develop a novel CRISPR/Cas9-based strategy for in situ directed evolution of mammalian proteins.
  • To identify novel fluorescent proteins with enhanced stability and functionality in specific cellular compartments.

Main Methods:

  • CRISPR/Cas9-based directed evolution.
  • In situ selection of functional protein variants within mammalian cells.
  • Fluorescent protein screening and characterization.

Main Results:

  • A new CRISPR/Cas9 strategy for directed protein evolution was established.
  • Functional mRuby3 variants were selected in lysosomes.
  • A novel fluorescent protein, mCRISPRed, was identified, exhibiting high stability and activity at low pH.

Conclusions:

  • The developed CRISPR/Cas9 method enables efficient in situ directed evolution of mammalian proteins.
  • mCRISPRed is a promising fluorescent reporter for low pH environments, such as lysosomes.
  • This work advances protein engineering tools for cellular and molecular biology research.