- Universal Loop assembly (uLoop): open, efficient, and species-agnostic DNA fabrication
- MoChlo: A versatile modular cloning toolbox for chloroplast biotechnology
- Golden Mutagenesis: An efficient multi-site saturation mutagenesis approach by Golden Gate cloning with automated primer design
- CyanoGate: A Golden Gate modular cloning suite for engineering cyanobacteria based on the plant MoClo syntax
- Birth of a photosynthetic chassis: a MoClo toolkit enabling synthetic biology in the microalga Chlamydomonas reinhardtii
- Loop Assembly: a simple and open system for recursive fabrication of DNA circuits
- AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach
- GoldenBraid 2.0: A Comprehensive DNA Assembly Framework for Plant Synthetic Biology
- A Golden Gate Modular Cloning Toolbox for Plants
- A versatile and robust Agrobacterium‐based gene stacking system generates high‐quality transgenic Arabidopsis plants
- A robust gene-stacking method utilizing yeast assembly for plant synthetic biology
Universal Loop assembly (uLoop): open, efficient, and species-agnostic DNA fabrication
Standardised Type IIS DNA assembly methods are becoming essential for biological engineering and research. Although a common syntax has been proposed to enable higher interoperability between DNA libraries, Golden Gate (GG) -based assembly systems remain specific to target organisms. Furthermore, these GG assembly systems become laborious and unnecessarily complicated beyond the assembly of 4 transcriptional units. Here, we describe 'universal Loop' (uLoop) assembly, a simple system based on Loop assembly that enables hierarchical fabrication of large DNA constructs (> 30 kb) for any organism of choice. uLoop comprises two sets of four plasmids that are iteratively used as odd and even levels to compile DNA elements in an exponential manner (4n-1). The elements required for transformation/maintenance in target organisms are also assembled as standardised parts, enabling customisation of host-specific plasmids. Thus, this species-agnostic method decouples efficiency of assembly from the stability of vectors in the target organism. As a proof-of-concept, we show the engineering of multi-gene expression vectors in diatoms, yeast, plants and bacteria. These resources will become available through the OpenMTA for unrestricted sharing and open-access.
Read the full preprint here.