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Related Experiment Video

Updated: Jun 3, 2026

CRISPR-based Shuttle Cloning: A High-throughput Cloning Method

CRISPR-based Shuttle Cloning: A High-throughput Cloning Method

Published on: June 13, 2025

A versatile and efficient high-throughput cloning tool for structural biology.

Eric R Geertsma¹, Raimund Dutzler

¹Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. e.geertsma@bioc.uzh.ch

|March 18, 2011

Summary

This summary is machine-generated.

Fragment Exchange (FX) cloning enables high-throughput generation of expression constructs for structural biology. This efficient method minimizes extra amino acids, speeding up expression screening for various systems.

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Area of Science:

Molecular Biology
Structural Biology
Biotechnology

Background:

High-throughput cloning is crucial for structural biology projects.
Existing methods have limitations in efficiency and sequence extension.

Purpose of the Study:

To introduce a novel cloning system, Fragment Exchange (FX) cloning.
To facilitate high-throughput generation of expression constructs.

Main Methods:

Utilizes a class IIS restriction enzyme and negative selection markers.
Combines features of recombination- and ligation-independent cloning.
Enables straightforward transfer of open reading frames into diverse expression vectors.

Main Results:

FX cloning is highly efficient and economical.
Minimizes extension of open reading frames with only a single extra amino acid.
Demonstrated robustness across prokaryotic and eukaryotic expression systems.

Conclusions:

FX cloning significantly accelerates the generation of expression constructs.
Facilitates broader expression screening for challenging structural biology projects.
Offers a robust and versatile alternative to traditional cloning methods.