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

Functional Editing of the OTC locus by Targeted Integration with Phenotype Correction and Restoration of Physiological Patterns of Expression.

Molecular therapy : the journal of the American Society of Gene Therapy·2026
Same author

ALS-FTD-linked CCNF<sup>S621G</sup> drives increased hippocampal astrocyte ramification and mitochondrial dysfunction and impairs motor neuron excitability.

Journal of neuroinflammation·2026
Same author

Renovating Neural Networks With Viral-Mediated Gene Transfer From A Tissue Contacting Matrix Mimic.

Small (Weinheim an der Bergstrasse, Germany)·2026
Same author

Wild-type and engineered adeno-associated viral vectors produce comparable opsin expression and light-evoked responses in rat skeletal muscle.

Molecular therapy. Methods & clinical development·2025
Same author

Genotypic, functional, and phenotypic characterization in CTNNB1 neurodevelopmental syndrome.

HGG advances·2025
Same author

High-throughput evaluation of cardiac-specific promoters for adeno-associated virus mediated cardiac gene therapy.

Gene therapy·2025

Related Experiment Video

Updated: Apr 24, 2026

Isolation of Next-Generation Gene Therapy Vectors through Engineering, Barcoding, and Screening of Adeno-Associated Virus AAV Capsid Variants
09:20

Isolation of Next-Generation Gene Therapy Vectors through Engineering, Barcoding, and Screening of Adeno-Associated Virus AAV Capsid Variants

Published on: October 18, 2022

5.1K

Selecting Novel Retino-tropic AAV Capsids by Directed Evolution.

Adrian Westhaus1,2, Leszek Lisowski3,4,5

  • 1Translational Vectorology Research Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Westmead, NSW, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|April 22, 2026
PubMed
Summary

This study details a comprehensive protocol for generating adeno-associated virus (AAV) libraries and selecting functional variants for gene therapy applications. The methods enable efficient AAV bioengineering for improved retinal transduction.

Keywords:
AAVCapsid library selectionDirected evolutionGene therapyViral vector production

More Related Videos

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

28.1K
Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors
09:12

Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors

Published on: April 5, 2024

5.1K

Related Experiment Videos

Last Updated: Apr 24, 2026

Isolation of Next-Generation Gene Therapy Vectors through Engineering, Barcoding, and Screening of Adeno-Associated Virus AAV Capsid Variants
09:20

Isolation of Next-Generation Gene Therapy Vectors through Engineering, Barcoding, and Screening of Adeno-Associated Virus AAV Capsid Variants

Published on: October 18, 2022

5.1K
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

28.1K
Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors
09:12

Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors

Published on: April 5, 2024

5.1K

Area of Science:

  • Molecular Biology
  • Gene Therapy
  • Biotechnology

Background:

  • Adeno-associated virus (AAV) vectors are crucial for gene therapy, particularly for retinal transduction.
  • Existing methods lack a complete, published protocol for directed evolution of AAV variants.
  • A detailed protocol is needed to guide library generation and selection of high-performing AAV candidates.

Purpose of the Study:

  • To provide a comprehensive, step-by-step protocol for generating AAV variant libraries.
  • To describe a transgene expression-driven selection platform for recovering functional AAV candidates.
  • To enable researchers to engineer novel AAV variants for enhanced gene delivery applications.

Main Methods:

  • Generation of a peptide insertion library into the AAV2 capsid's variable region VIII.
  • Utilizing a highly predictive transgene expression-driven selection platform for candidate recovery.
  • Adaptable protocol applicable to various AAV serotypes and surface loops, independent of model systems.

Main Results:

  • The protocol facilitates the creation of diverse AAV libraries.
  • The selection platform effectively identifies functional AAV variants with improved transduction capabilities.
  • The described methods are accessible and do not require specialized equipment.

Conclusions:

  • This detailed protocol empowers researchers in AAV bioengineering and gene therapy.
  • The methods support the development of novel AAV variants for enhanced retinal gene delivery.
  • The protocol is a valuable resource for advancing AAV-based therapeutic strategies.