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Related Concept Videos

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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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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Riboswitches01:56

Riboswitches

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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
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Bacterial Transcription01:53

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RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
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Transcription Attenuation in Prokaryotes02:42

Transcription Attenuation in Prokaryotes

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Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure.  Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
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In-vitro Mutagenesis01:16

In-vitro Mutagenesis

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To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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Cell-free Protein Expression Using the Rapidly Growing Bacterium Vibrio natriegens
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Mobile-CRISPRi as a powerful tool for modulating Vibrio gene expression.

Logan Geyman, Madeline Tanner, Natalia Rosario-Melendez

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    Mobile-CRISPRi enables robust gene knockdown in diverse Vibrio species. This system precisely targets genes to study bacterial functions like bioluminescence and growth.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • CRISPR interference (CRISPRi) is a powerful gene knockdown technique utilizing a deactivated Cas9 protein (dCas9) and a guide RNA (sgRNA) to block gene transcription.
    • Investigating gene function in diverse bacterial species, particularly those with medical or ecological significance like Vibrio, is crucial for understanding their biology.

    Approach:

    • The Mobile-CRISPRi system employs modular vectors for integrating IPTG-inducible dcas9 and sgRNA genes into bacterial genomes via Tn7 transposition.
    • This study validates the Mobile-CRISPRi system's functionality and specificity across multiple Vibrio species, including V. cholerae, V. fischeri, V. vulnificus, V. parahaemolyticus, and V. campbellii.

    Key Points:

    • The Mobile-CRISPRi system demonstrates robust and specific gene knockdown in a range of Vibrio species.
    • Efficacy was shown by targeting essential and non-essential genes to modulate phenotypes such as bioluminescence, quorum sensing, cell division, and growth arrest.
    • Successful gene targeting was achieved for phenotypes including bioluminescence, quorum sensing, cell division, and growth arrest.

    Conclusions:

    • Mobile-CRISPRi provides a versatile tool for functional genomics in Vibrio species.
    • This system is expected to accelerate the systematic investigation of gene function and essentiality in Vibrio, aiding research in diverse environments and behaviors.