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Updated: May 16, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

Highly replicated experiments studying complex genotypes using nested DNA barcodes.

Molly Monge^1,2, Simone M Giovanetti¹, Apoorva Ravishankar^1,3

¹Center for Genomics and Data Science Research, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.

Biorxiv : the Preprint Server for Biology

|April 1, 2025

Summary

This summary is machine-generated.

We developed a novel barcode method to track numerous biological replicates of complex genetic modifications. This approach enables robust detection of subtle phenotypic differences in high-throughput genetic screens.

Keywords:

DNA barcodes NER factor I complex cerevisiae yeast

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Competitive Genomic Screens of Barcoded Yeast Libraries

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Published on: August 11, 2011

Area of Science:

Genetics
Molecular Biology
Biotechnology

Background:

Studying genetic modifications, especially combinatorial genotypes, presents challenges in sample replication and tracking.
Increased genotype complexity escalates the difficulty of generating and managing independent biological replicates.

Purpose of the Study:

To develop a scalable, barcode-based method for tracking large numbers of independent replicates of combinatorial genotypes.
To enable robust detection of subtle phenotypic differences in pooled genetic libraries.
To provide a framework for exploring complex genotype-phenotype relationships.

Main Methods:

Utilized a nested serial cloning process to create a plasmid library of combinatorial genotypes with DNA barcodes.
Combined gene variants with associated DNA barcodes, creating plasmids with multiple gene variants and a combined barcode.
Employed next-generation sequencing to analyze the pooled barcodes, allowing study of the entire population in a single flask.

Main Results:

Demonstrated the ability to track numerous independent replicates of complex genotypes in a pooled format.
Identified superior growth in DNA-damaging conditions for cells expressing all three yeast NEF-1 subunits.
Assessed method sensitivity through simulated downsampling of barcodes.

Conclusions:

The NICR barcode method is effective for high-throughput combinatorial genetic screens.
The developed framework offers a scalable solution for investigating complex genotype-phenotype relationships.
This approach facilitates robust detection of subtle phenotypic differences.