Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Dissecting conserved molecular mechanisms of biological toxin activity through CRISPR screening.

Biochemical Society transactions·2026
Same author

Correction: The cytotoxicity of gomesin peptides is mediated by the glycosphingolipid pathway and lipid-cholesterol interactions.

Cell death discovery·2026
Same author

Identification of the MRTFA/SRF pathway as a critical regulator of quiescence and chemotherapy resistance in cancer.

Cancer letters·2026
Same author

Histone methyltransferase PRDM9 promotes survival of drug-tolerant persister cells in glioblastoma.

Nature communications·2025
Same author

Unravelling the magnitude and drivers of PFAS trophic magnification: a meta-analysis.

Nature communications·2025
Same author

The cytotoxicity of gomesin peptides is mediated by the glycosphingolipid pathway and lipid-cholesterol interactions.

Cell death discovery·2025

Related Experiment Video

Updated: May 12, 2025

Engineering and Evolution of Synthetic Adeno-Associated Virus AAV Gene Therapy Vectors via DNA Family Shuffling
21:55

Engineering and Evolution of Synthetic Adeno-Associated Virus AAV Gene Therapy Vectors via DNA Family Shuffling

Published on: April 2, 2012

28.3K

A chimeric viral platform for directed evolution in mammalian cells.

Alexander J Cole1, Christopher E Denes2, Cesar L Moreno2

  • 1Centenary Institute and Faculty of Medicine and Health, Charles Perkins Centre, The University of Sydney, Sydney, NSW, Australia.

Nature Communications
|May 7, 2025
PubMed
Summary
This summary is machine-generated.

We developed a novel mammalian directed evolution platform, PROTein Evolution Using Selection (PROTEUS), enabling stable and efficient biomolecule engineering. This system enhances gene regulation tools and nanobody evolution in mammalian cells.

More Related Videos

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs
11:21

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

Published on: September 10, 2017

9.4K
Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity
10:39

Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity

Published on: June 25, 2015

12.5K

Related Experiment Videos

Last Updated: May 12, 2025

Engineering and Evolution of Synthetic Adeno-Associated Virus AAV Gene Therapy Vectors via DNA Family Shuffling
21:55

Engineering and Evolution of Synthetic Adeno-Associated Virus AAV Gene Therapy Vectors via DNA Family Shuffling

Published on: April 2, 2012

28.3K
Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs
11:21

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

Published on: September 10, 2017

9.4K
Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity
10:39

Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity

Published on: June 25, 2015

12.5K

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Synthetic Biology

Background:

  • Directed evolution is crucial for developing novel biomolecules but faces challenges in mammalian systems.
  • Existing prokaryotic and yeast-based platforms are not ideal for all mammalian cell applications.
  • Stable and efficient mammalian-based directed evolution systems are needed.

Purpose of the Study:

  • To develop a stable and efficient platform for directed evolution in mammalian cells.
  • To demonstrate the utility of the platform for engineering gene regulation tools and intracellular nanobodies.
  • To overcome limitations of current directed evolution systems in mammalian contexts.

Main Methods:

  • Development of the PROTein Evolution Using Selection (PROTEUS) platform using chimeric virus-like vesicles.
  • Application of PROTEUS for altering doxycycline responsiveness of tetracycline-controlled transactivators.
  • Utilizing PROTEUS for intracellular nanobody evolution, specifically targeting a DNA damage-responsive anti-p53 nanobody.

Main Results:

  • The PROTEUS platform enables extended mammalian directed evolution campaigns without compromising system integrity.
  • A more sensitive TetON-4G gene regulation tool was generated with mammalian-specific adaptations.
  • Successful evolution of a DNA damage-responsive anti-p53 nanobody was achieved using the PROTEUS platform.

Conclusions:

  • PROTEUS is an efficient and stable platform for directed evolution of biomolecules within mammalian cells.
  • The platform facilitates the generation of novel tools for gene regulation and protein engineering in mammalian systems.
  • PROTEUS represents a significant advancement for mammalian-based biotechnology and synthetic biology.