Error correction of microchip synthesized genes using Surveyor nuclease

Area of Science:

• Molecular Biology
• Synthetic Biology
• Biotechnology

Background:

• High-throughput gene synthesis is crucial for biotechnology but limited by errors.
• Current error correction methods are costly and time-consuming.

Purpose of the Study:

• To develop an efficient and economical method for correcting errors in synthetic genes.
• To enhance the fidelity of gene synthesis technologies.

Main Methods:

• An error correction reaction (ECR) utilizing Surveyor endonuclease was developed.
• ECR involves re-annealing synthetic genes to reveal mismatches, followed by enzymatic excision of errors.
• Overlap extension polymerase chain reaction (OE-PCR) was used to reassemble corrected gene fragments.

Main Results:

• The ECR method effectively removes errors from synthetic genes.
• Iterating the ECR process increased gene synthesis fidelity.
• Two iterations of ECR reduced errors by over 16-fold, achieving an error rate of approximately 1 in 8700 base pairs.

Conclusions:

• ECR is a highly effective strategy for improving the accuracy of synthetic genes.
• This method offers a cost-effective and high-throughput solution for gene synthesis error correction.
• ECR has the potential to significantly advance synthetic biology applications.