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  1. Article ; Online: CS-Cells: A CRISPR-Cas12 DNA Device to Generate Chromosome-Shredded Cells for Efficient and Safe Molecular Biomanufacturing.

    Pantoja Angles, Aarón / Ali, Zahir / Mahfouz, Magdy

    ACS synthetic biology

    2022  Volume 11, Issue 1, Page(s) 430–440

    Abstract: Synthetic biology holds great promise for translating ideas into products to address the grand challenges facing humanity. Molecular biomanufacturing is an emerging technology that facilitates the production of key products of value, including ... ...

    Abstract Synthetic biology holds great promise for translating ideas into products to address the grand challenges facing humanity. Molecular biomanufacturing is an emerging technology that facilitates the production of key products of value, including therapeutics and select chemical compounds. Current biomanufacturing technologies require improvements to overcome limiting factors, including efficient production, cost, and safe release; therefore, developing optimum chassis for biomolecular manufacturing is of great interest for enabling diverse synthetic biology applications. Here, we harnessed the power of the CRISPR-Cas12 system to design, build, and test a DNA device for genome shredding, which fragments the native genome to enable the conversion of bacterial cells into nonreplicative, biosynthetically active, and programmable molecular biomanufacturing chassis. As a proof of concept, we demonstrated the efficient production of green fluorescent protein and violacein, an antimicrobial and antitumorigenic compound. Our CRISPR-Cas12-based chromosome-shredder DNA device has built-in biocontainment features providing a roadmap for the conversion of any bacterial cell into a chromosome-shredded chassis amenable to high-efficiency molecular biomanufacturing, thereby enabling exciting and diverse biotechnological applications.
    MeSH term(s) Biotechnology ; CRISPR-Cas Systems/genetics ; Chromosomes ; DNA/genetics ; Synthetic Biology
    Chemical Substances DNA (9007-49-2)
    Language English
    Publishing date 2022-01-03
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 2161-5063
    ISSN (online) 2161-5063
    DOI 10.1021/acssynbio.1c00516
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Microbial Biocontainment Systems for Clinical, Agricultural, and Industrial Applications.

    Pantoja Angles, Aaron / Valle-Pérez, Alexander U / Hauser, Charlotte / Mahfouz, Magdy M

    Frontiers in bioengineering and biotechnology

    2022  Volume 10, Page(s) 830200

    Abstract: Many applications of synthetic biology require biological systems in engineered microbes to be delivered into diverse environments, such as ... ...

    Abstract Many applications of synthetic biology require biological systems in engineered microbes to be delivered into diverse environments, such as for
    Language English
    Publishing date 2022-02-02
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2719493-0
    ISSN 2296-4185
    ISSN 2296-4185
    DOI 10.3389/fbioe.2022.830200
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Efficient in planta production of amidated antimicrobial peptides that are active against drug-resistant ESKAPE pathogens.

    Chaudhary, Shahid / Ali, Zahir / Tehseen, Muhammad / Haney, Evan F / Pantoja-Angles, Aarón / Alshehri, Salwa / Wang, Tiannyu / Clancy, Gerard J / Ayach, Maya / Hauser, Charlotte / Hong, Pei-Ying / Hamdan, Samir M / Hancock, Robert E W / Mahfouz, Magdy

    Nature communications

    2023  Volume 14, Issue 1, Page(s) 1464

    Abstract: Antimicrobial peptides (AMPs) are promising next-generation antibiotics that can be used to combat drug-resistant pathogens. However, the high cost involved in AMP synthesis and their short plasma half-life render their clinical translation a challenge. ... ...

    Abstract Antimicrobial peptides (AMPs) are promising next-generation antibiotics that can be used to combat drug-resistant pathogens. However, the high cost involved in AMP synthesis and their short plasma half-life render their clinical translation a challenge. To address these shortcomings, we report efficient production of bioactive amidated AMPs by transient expression of glycine-extended AMPs in Nicotiana benthamiana line expressing the mammalian enzyme peptidylglycine α-amidating mono-oxygenase (PAM). Cationic AMPs accumulate to substantial levels in PAM transgenic plants compare to nontransgenic N. benthamiana. Moreover, AMPs purified from plants exhibit robust killing activity against six highly virulent and antibiotic resistant ESKAPE pathogens, prevent their biofilm formation, analogous to their synthetic counterparts and synergize with antibiotics. We also perform a base case techno-economic analysis of our platform, demonstrating the potential economic advantages and scalability for industrial use. Taken together, our experimental data and techno-economic analysis demonstrate the potential use of plant chassis for large-scale production of clinical-grade AMPs.
    MeSH term(s) Animals ; Anti-Bacterial Agents/biosynthesis ; Anti-Bacterial Agents/pharmacology ; Antimicrobial Cationic Peptides/biosynthesis ; Antimicrobial Cationic Peptides/pharmacology ; Antimicrobial Peptides/biosynthesis ; Mammals ; Plants ; Nicotiana/chemistry ; Nicotiana/genetics ; Drug Resistance, Bacterial/drug effects
    Chemical Substances Anti-Bacterial Agents ; Antimicrobial Cationic Peptides ; Antimicrobial Peptides
    Language English
    Publishing date 2023-03-16
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2553671-0
    ISSN 2041-1723 ; 2041-1723
    ISSN (online) 2041-1723
    ISSN 2041-1723
    DOI 10.1038/s41467-023-37003-z
    Database MEDical Literature Analysis and Retrieval System OnLINE

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