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

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

5.9K
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...
5.9K
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

6.6K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
6.6K

You might also read

Related Articles

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

Sort by
Same author

FN-silk membrane enables alveologenesis processes and self-organization of the H441 epithelial cell line into native-like alveolar morphology.

Scientific reports·2026
Same author

Recombinant spider silk membranes promote human renal epithelial differentiation and function.

Biofabrication·2026
Same author

Recombinant spider silk functionalized with a CD40 agonist shows improved capability to activate human B cells in vitro - A novel module for cancer immunotherapy.

International journal of biological macromolecules·2025
Same author

3D Culture in Functionalized FN-Silk Networks Facilitate Proliferation, Differentiation and Phenotypic Stability of Mature Human Primary Cells and Stem Cells.

Biotechnology and bioengineering·2025
Same author

A Fibronectin (FN)-Silk 3D Cell Culture Model as a Screening Tool for Repurposed Antifibrotic Drug Candidates for Endometriosis.

Small (Weinheim an der Bergstrasse, Germany)·2025
Same author

Site-Specific Immobilization Boosts the Performance of a Galectin-1 Biosensor.

Bioconjugate chemistry·2024

Related Experiment Video

Updated: May 26, 2025

Material Formation of Recombinant Spider Silks through Aqueous Solvation using Heat and Pressure
10:26

Material Formation of Recombinant Spider Silks through Aqueous Solvation using Heat and Pressure

Published on: May 6, 2019

5.1K

Site-Specific Functionalization of Recombinant Spider Silk Using Enzymatic Sortase Coupling.

Rajeev Pasupuleti1, Ronnie Jansson1, Ida Isacsson1

  • 1KTH Royal Institute of Technology, School of Engineering Sciences in Chemistry, Biotechnology and Health, Department of Protein Science, AlbaNova University Center, Roslagstullsbacken 21, SE-106 91 Stockholm, Sweden.

ACS Omega
|February 24, 2025
PubMed
Summary

Researchers developed a site-specific method to attach proteins to spider silk biomaterials using sortase A enzyme. This technique efficiently functionalizes silk coatings and fibers, creating advanced biomaterials for research.

More Related Videos

Synthetic Spider Silk Production on a Laboratory Scale
13:36

Synthetic Spider Silk Production on a Laboratory Scale

Published on: July 18, 2012

26.7K
Author Spotlight: Enhancing In Vitro Cell Culture Models with Recombinant Functionalized Spider Silk Membranes
06:17

Author Spotlight: Enhancing In Vitro Cell Culture Models with Recombinant Functionalized Spider Silk Membranes

Published on: November 1, 2024

910

Related Experiment Videos

Last Updated: May 26, 2025

Material Formation of Recombinant Spider Silks through Aqueous Solvation using Heat and Pressure
10:26

Material Formation of Recombinant Spider Silks through Aqueous Solvation using Heat and Pressure

Published on: May 6, 2019

5.1K
Synthetic Spider Silk Production on a Laboratory Scale
13:36

Synthetic Spider Silk Production on a Laboratory Scale

Published on: July 18, 2012

26.7K
Author Spotlight: Enhancing In Vitro Cell Culture Models with Recombinant Functionalized Spider Silk Membranes
06:17

Author Spotlight: Enhancing In Vitro Cell Culture Models with Recombinant Functionalized Spider Silk Membranes

Published on: November 1, 2024

910

Area of Science:

  • Biomaterials Science
  • Protein Engineering
  • Biotechnology

Background:

  • Recombinant spider silk proteins like FN-4RepCT offer versatile formats (coatings, fibers) for biomedical applications.
  • Site-specific functionalization is crucial for developing next-generation biomaterials with tailored properties.
  • Enzymatic coupling methods provide mild and efficient strategies for biomaterial modification.

Purpose of the Study:

  • To report a sortase A enzymatic coupling method for site-specific functionalization of FN-4RepCT spider silk formats.
  • To demonstrate the attachment of diverse functional proteins (Z-domain, scFvCD38, Sal-1) to spider silk.
  • To compare the efficiency of sortase coupling using different spider silk variants (G-silk vs. G5-silk).

Main Methods:

  • Utilized sortase A enzymatic coupling with an LPETGG tag on target proteins.
  • Functionalized various FN-4RepCT formats including coatings, nanowires, and fibers.
  • Assessed coupling efficiency and retained protein activity using gel electrophoresis, fluorescence microscopy, SPR, and biochemical assays.

Main Results:

  • The sortase A method successfully enabled site-specific functionalization of FN-4RepCT with Z-domain, scFvCD38, and Sal-1.
  • FN-4RepCT with five N-terminal glycines (G5-silk) demonstrated higher sortase coupling efficiency compared to G-silk.
  • All coupled proteins retained their specific biological activity after attachment to the silk biomaterials.

Conclusions:

  • Sortase coupling is a mild and efficient strategy for site-specifically functionalizing various spider silk formats.
  • This approach allows the attachment of both small and large functional proteins, expanding biomaterial capabilities.
  • The developed method facilitates the creation of advanced, customized biomaterials for biomedical research and applications.