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Assembly of Customized TAL Effectors Through Advanced ULtiMATE System.
Junjiao Yang1, Shengjie Guo1, Pengfei Yuan1
1Biodynamic Optical Imaging Center (BIOPIC), State Key Laboratory of Protein and Plant Gene Research, College of Life Sciences, Peking University, Beijing, 100871, China.
Methods in Molecular Biology (Clifton, N.J.)
|October 8, 2015
Summary
The ULtiMATE system enables rapid, efficient construction of Transcription Activator-Like Effectors (TALEs) using USER fusion and a tetramer template archive. This advanced method streamlines the assembly of TALEs, TALENs, and other TALE-based constructs in just half a day.
Area of Science:
- Molecular Biology
- Synthetic Biology
- Gene Editing Technologies
Background:
- Transcription Activator-Like Effectors (TALEs) are crucial tools for gene targeting.
- Existing methods for constructing TALEs can be time-consuming and complex.
Purpose of the Study:
- To develop an advanced system for highly efficient and rapid construction of TALEs.
- To create a versatile platform for assembling various TALE-based constructs, including TALENs.
Main Methods:
- Utilized the USER (Uracil-Specific Excision and Re-ligation) fusion technique.
- Employed an archive of 512 distinct tetramer DNA templates for TALE assembly.
- Developed the ULtiMATE (USER-based Ligation-Mediated Assembly of TAL Effector) system.
Main Results:
- Achieved highly efficient construction of TALE DNA constructs.
- Reduced the assembly time for any given TALE construct to approximately half a day.
- Demonstrated suitability for large-scale assembly of TALENs and other TALE-based systems.
Conclusions:
- The ULtiMATE system offers a significant advancement in TALE construction efficiency and speed.
- This method provides a streamlined approach for researchers working with TALE-based gene editing tools.
- The system is adaptable for various applications requiring TALE protein engineering.
