Abstract
The present invention provides methods and systems for modifying intron sequences to enable their use in modular cloning (MoClo) systems while preserving their ability to enhance gene accumulation. The invention addresses a critical challenge in molecular cloning where beneficial introns contain restriction enzyme recognition sites that interfere with MoClo assembly. The methods include computational approaches for identifying minimal sequence modifications needed to eliminate restriction sites while maintaining intron functionality, experimental validation protocols for confirming preserved enhancement properties, and high-throughput screening systems for rapidly assessing multiple intron variants. In particular embodiments, the invention demonstrates successful modification of introns for enhanced accumulation of bovine casein genes in transgenic soybean, enabling efficient assembly of complex multi-gene constructs while maintaining optimal protein production. The invention provides a systematic and reproducible approach to intron domestication that can be applied across different gene systems and host organisms. Alternative embodiments include modified restriction enzyme strategies and methods for generating validated libraries of MoClo-compatible introns with documented enhancement effects.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
Recommended Citation
Tobin, Cory, "Methods and Systems for Modifying Intron Sequences to Remove Restriction Sites While Preserving Gene Accumulation Enhancement in Transgenic Plants", Technical Disclosure Commons, (February 09, 2026)
https://www.tdcommons.org/dpubs_series/9302