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Title: Chapter 2: Trackable Multiplex Recombineering (TRMR) and Next-Generation Genome Design Technologies: Modifying Gene Expression in E. coli by Inserting Synthetic DNA Cassettes and Molecular Barcodes

Abstract

This chapter describes the trackable multiplex recombineering (TRMR) and tunable trackable multiplex recombineering (T2RMR) techniques, which not only make the multiplexing of recombineering possible in Escherichia coli but also provide the ability to track the engineered genetic changes accurately. TRMR and T2RMR have only been used for modifying the expression level of genes. Screening and selection of TRMR and T2RMR clones can be carried out using either liquid or solid media with any chemical compound that modifies growth or confers a phenotype that can be detected by a high-throughput assay. In order to engineer a genome using TRMR or T2RMR, a synthetic DNA (synDNA) cassette is created that encodes for a genetic feature and a molecular barcode that is used to track each feature. These synDNA cassettes are then introduced in parallel into cell populations via recombineering.

Authors:
 [1]; ORCiD logo [1];  [2];  [2]
  1. National Renewable Energy Laboratory (NREL), Golden, CO (United States)
  2. University of Colorado
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE)
OSTI Identifier:
1465638
Report Number(s):
NREL/CH-2700-72219
DOE Contract Number:  
AC36-08GO28308
Resource Type:
Book
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; E. coli; gene expression; genetic feature; molecular barcodes; next-generation genome design technologies; synthetic DNA cassette; tunable trackable multiplex recombineering

Citation Formats

Freed, Emily, Eckert, Carrie A, Pines, Gur, and Gill, Ryan T. Chapter 2: Trackable Multiplex Recombineering (TRMR) and Next-Generation Genome Design Technologies: Modifying Gene Expression in E. coli by Inserting Synthetic DNA Cassettes and Molecular Barcodes. United States: N. p., 2018. Web. doi:10.1002/9783527688104.ch2.
Freed, Emily, Eckert, Carrie A, Pines, Gur, & Gill, Ryan T. Chapter 2: Trackable Multiplex Recombineering (TRMR) and Next-Generation Genome Design Technologies: Modifying Gene Expression in E. coli by Inserting Synthetic DNA Cassettes and Molecular Barcodes. United States. doi:10.1002/9783527688104.ch2.
Freed, Emily, Eckert, Carrie A, Pines, Gur, and Gill, Ryan T. Sat . "Chapter 2: Trackable Multiplex Recombineering (TRMR) and Next-Generation Genome Design Technologies: Modifying Gene Expression in E. coli by Inserting Synthetic DNA Cassettes and Molecular Barcodes". United States. doi:10.1002/9783527688104.ch2.
@article{osti_1465638,
title = {Chapter 2: Trackable Multiplex Recombineering (TRMR) and Next-Generation Genome Design Technologies: Modifying Gene Expression in E. coli by Inserting Synthetic DNA Cassettes and Molecular Barcodes},
author = {Freed, Emily and Eckert, Carrie A and Pines, Gur and Gill, Ryan T.},
abstractNote = {This chapter describes the trackable multiplex recombineering (TRMR) and tunable trackable multiplex recombineering (T2RMR) techniques, which not only make the multiplexing of recombineering possible in Escherichia coli but also provide the ability to track the engineered genetic changes accurately. TRMR and T2RMR have only been used for modifying the expression level of genes. Screening and selection of TRMR and T2RMR clones can be carried out using either liquid or solid media with any chemical compound that modifies growth or confers a phenotype that can be detected by a high-throughput assay. In order to engineer a genome using TRMR or T2RMR, a synthetic DNA (synDNA) cassette is created that encodes for a genetic feature and a molecular barcode that is used to track each feature. These synDNA cassettes are then introduced in parallel into cell populations via recombineering.},
doi = {10.1002/9783527688104.ch2},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2018},
month = {3}
}

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