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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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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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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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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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Cooperative Binding of Transcription Regulators02:13

Cooperative Binding of Transcription Regulators

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Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
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Related Experiment Video

Updated: Dec 11, 2025

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9
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Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

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Conditional Recruitment to a DNA-Bound CRISPR-Cas Complex Using a Colocalization-Dependent Protein Switch.

Robin L Kirkpatrick, Kieran Lewis, Robert A Langan

    ACS Synthetic Biology
    |August 21, 2020
    PubMed
    Summary

    Researchers developed a novel synthetic switch (Co-LOCKR) that activates upon binding to specific DNA target sites. This DNA-triggered system enables precise spatial control of biochemical functions within the genome.

    Keywords:
    CRISPR−CasCo-LOCKRgenetic circuitprotein switch

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    Area of Science:

    • Synthetic Biology
    • Molecular Biology
    • Genetics

    Background:

    • Precise spatial control of genomic functions is essential for advanced synthetic biology applications.
    • Existing methods lack specificity in targeting and activating functions at defined genomic loci.

    Purpose of the Study:

    • To engineer a DNA-triggered synthetic switch for site-specific genomic function activation.
    • To develop a platform for sophisticated genetic control using CRISPR-Cas technology.

    Main Methods:

    • Engineered a de novo protein switch (Co-LOCKR) activated by DNA binding.
    • Utilized two CRISPR-Cas complexes to colocalize switch components to adjacent genomic sites.
    • Prototyped the system in yeast to demonstrate DNA-triggered activation and reporter gene expression.

    Main Results:

    • Demonstrated that DNA binding induces a conformational change in the Co-LOCKR switch, transitioning it to an active state.
    • Showcased successful recruitment of a transcription factor upon switch activation.
    • Confirmed expression of a downstream reporter gene, validating the system's functionality.

    Conclusions:

    • The DNA-triggered Co-LOCKR switch offers a novel platform for precise spatial control of biochemical functions.
    • This system has potential applications in epigenetic regulation, biological imaging, and genetic logic circuits.
    • The technology enables sophisticated genomic functions to be executed exclusively at designated target sites.