Related Experiment Video
Updated: Jan 26, 2026
06:28
Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit
Published on: September 2, 2025
776
iSeq 2.0: A Modular and Interchangeable Toolkit for Interaction Screening in Yeast.
Xianan Liu1, Zhimin Liu1, Adam K Dziulko1
1Department of Biochemistry, Stony Brook University, Stony Brook, NY 11794-5215, USA; Laufer Center for Physical and Quantitative Biology, Stony Brook University, Stony Brook, NY 11794-5252, USA.
Cell Systems
|April 8, 2019
Summary
We created a flexible toolkit for combinatorial screening in yeast. This platform, iSeq 2.0, uses DNA barcoding to enable large-scale screening of genetic element combinations via short-read sequencing.
Area of Science:
- Synthetic Biology
- Genomics
- Molecular Biology
Background:
- Combinatorial screening is crucial for understanding gene function and complex biological systems.
- Existing methods for generating and analyzing large-scale combinatorial libraries in yeast have limitations in flexibility and scalability.
Purpose of the Study:
- To develop a versatile and scalable toolkit for combinatorial screening in Saccharomyces cerevisiae.
- To enable the generation of large, uniquely barcoded cell libraries for high-throughput genetic analysis.
Main Methods:
- Developed the interaction sequencing (iSeq 2.0) platform, incorporating genomic landing pads for sequential DNA element integration.
- Utilized 15 barcoded plasmid libraries with split selectable markers and an array of double-barcoder strains.
- Enabled compatibility with various DNA elements, including integrating plasmids, genomic modifications, and genetic backgrounds.
Main Results:
- The iSeq 2.0 platform generates large libraries of uniquely barcoded cells, each representing a specific DNA element combination.
- Demonstrated modularity and interchangeability of DNA element libraries, allowing for flexible combinatorial designs.
- Showcased the capability to perform numerous new combinatorial screens using short-read sequencing.
Conclusions:
- The iSeq 2.0 toolkit provides a flexible and powerful platform for advancing combinatorial genetic screening in yeast.
- This technology facilitates high-throughput analysis of DNA element interactions and complex genetic architectures.
- iSeq 2.0 significantly expands the possibilities for yeast-based functional genomics and synthetic biology applications.
Related Concept Videos
01:57Protein Complexes with Interchangeable Parts
2.9K
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
2.9K
01:57Protein Complexes with Interchangeable Parts
2.1K
2.1K
01:28Yeast Signaling
17.2K
Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes (cells with a nucleus). Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule released from a yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases to activate or inactivate transcription factors that further regulate gene expression. Many of the yeast intracellular signaling cascades have similar...
17.2K
01:28Drug Products: Biologics, Biosimilars and Interchangeables
242
Body:Biologics, derived from living sources such as humans, animals, or microorganisms, represent a significant category of pharmaceuticals. These complex molecules, developed through advanced biotechnological methods or purified from natural sources, include essential medical treatments like insulin and growth hormones. The complexity of biologics arises from their large molecular structures and the intricate processes required for their production, making them distinct from conventional...
242
02:39Predator-Prey Interactions
21.2K
Predators consume prey for energy. Predators that acquire prey and prey that avoid predation both increase their chances of survival and reproduction (i.e., fitness). Routine predator-prey interactions elicit mutual adaptations that improve predator offenses, such as claws, teeth, and speed, as well as prey defenses, including crypsis, aposematism, and mimicry. Thus, predator-prey interactions resemble an evolutionary arms race.
21.2K
02:46Genetic Screens
5.6K
Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
5.6K