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Precise Phage Mutagenesis with NgTET-Assisted CRISPR-Cas Systems
Published on: October 14, 2025
M13 bacteriophage display framework that allows sortase-mediated modification of surface-accessible phage proteins
Gaelen T Hess1, Juan J Cragnolini, Maximilian W Popp
1The David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Bioconjugate Chemistry
|July 5, 2012
Summary
Bacterial sortases enable precise M13 bacteriophage capsid modification with diverse molecules and proteins, significantly improving display efficiency for materials science and biological applications.
Area of Science:
- Biotechnology
- Molecular Biology
- Materials Science
Background:
- M13 bacteriophage is a versatile platform for biological and materials science applications.
- Current methods for M13 functionalization have limitations in efficiency and scope.
- Site-specific modification of phage capsid proteins is crucial for advanced applications.
Purpose of the Study:
- To develop a novel method for site-specific functionalization of M13 bacteriophage capsid proteins.
- To enhance the display efficiency of various moieties onto M13 phage particles.
- To demonstrate the utility of sortase-mediated conjugation for M13 phage engineering.
Main Methods:
- Utilizing bacterial sortases for site-specific conjugation of molecules and proteins to M13 phage capsid proteins (pIII, pIX, pVIII).
- Employing orthogonal sortases for simultaneous targeting of multiple capsid proteins.
- Quantifying the efficiency of moiety display using sortase-based reactions.
Main Results:
- Achieved site-specific functionalization of pIII, pIX, and pVIII with diverse entities, including small molecules and proteins.
- Demonstrated significantly higher yields compared to traditional phage display methods.
- Reported a 100-fold increase in the efficiency of Green Fluorescent Protein (GFP) display on pVIII.
- Successfully achieved simultaneous labeling of two distinct capsid proteins with high specificity.
Conclusions:
- Sortase-mediated conjugation provides a simple, effective, and highly efficient method for M13 bacteriophage functionalization.
- This approach expands the capabilities of M13 phage as a tool in materials science and biotechnology.
- The enhanced display efficiency opens new avenues for constructing complex phage-based structures and applications.
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