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Updated: Jul 4, 2026

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

Rapid and efficient gene splicing using megaprimer-based protocol.

Ji-Ren Chen¹, Jing-Jing Lü, Hua-Fang Wang

¹College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, People's Republic of China.

Molecular Biotechnology

|June 25, 2008

Summary

This summary is machine-generated.

This study presents an improved megaprimer PCR method for efficiently splicing gene fragments. The modified technique enhances specificity and applicability for creating chimeric genes without restriction sites.

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Published on: January 8, 2015

Area of Science:

Molecular Biology
Biotechnology

Background:

Megaprimer PCR is common for site-directed mutagenesis but underutilized for gene splicing.
Existing gene splicing methods often rely on restriction sites, limiting flexibility.

Purpose of the Study:

To describe a modified megaprimer PCR method for efficient gene splicing.
To elucidate the mechanism and key factors of megaprimer-based gene ligation.
To provide a protocol for creating and amplifying ligated chimeric gene segments.

Main Methods:

Modification of the megaprimer PCR technique.
Utilizing denatured megaprimer strands as both template and primer.
Application of a common PCR program for ligated chimeric gene amplification.

Main Results:

Efficient creation and amplification of specific ligated chimeric gene segments.
Elucidation of the mechanism involving megaprimer strand regeneration and ligation.
Successful application to large chimeric gene products with increased specificity and efficiency.

Conclusions:

The improved megaprimer PCR protocol offers a simple and broadly applicable method for gene splicing.
This technique enables seamless ligation of gene fragments without dependence on restriction sites.
The findings provide a deeper understanding of megaprimer PCR mechanisms for gene manipulation.