Rewriting the blueprint of life by synthetic genomics and genome engineering.

Sivaprakash Ramalingam,Narayana Annaluru,Srinivasan Chandrasegaran

doi:10.1186/s13059-015-0689-y

Sivaprakash Ramalingam, Narayana Annaluru + Show 1 more

Open Access

https://doi.org/10.1186/s13059-015-0689-y

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Journal: Genome biology	Publication Date: Jun 16, 2015
Citations: 80	License type: CC BY 4.0

Affiliation: Johns Hopkins University

Abstract

Advances in DNA synthesis and assembly methods over the past decade have made it possible to construct genome-size fragments from oligonucleotides. Early work focused on synthesis of small viral genomes, followed by hierarchical synthesis of wild-type bacterial genomes and subsequently on transplantation of synthesized bacterial genomes into closely related recipient strains. More recently, a synthetic designer version of yeast Saccharomyces cerevisiae chromosome III has been generated, with numerous changes from the wild-type sequence without having an impact on cell fitness and phenotype, suggesting plasticity of the yeast genome. A project to generate the first synthetic yeast genome - the Sc2.0 Project - is currently underway.

Highlights

Biology is undergoing a rapid transition from the age of deciphering DNA sequence information of the genomes of biological species to the age of synthetic genomes
Genome editing approaches for genome-wide scale alteration that are not based on total synthesis of the genome are being pursued and have proved powerful; for example, in the production of a reduced-size genome version of Escherichia coli [4] and engineering of bacterial genomes to include many different changes simultaneously [8]
The hierarchical synthesis of the M. genitalium genome was done in three steps: (1) overlapping 5–7 kbp DNA fragments were assembled from chemically synthesized oligonucleotides; (2) the 5–7 kbp fragments were joined by in vitro recombination to yield intermediate 24 kbp, 72 kbp and 144 kbp fragments that were cloned into bacterial artificial chromosomes in E. coli; (3) the complete synthetic genome was assembled by homologous recombination in the yeast S. cerevisiae

Summary

Introduction

Biology is undergoing a rapid transition from the age of deciphering DNA sequence information of the genomes of biological species to the age of synthetic genomes. The authors replaced 11,515 bp of the 5′ part of the 39,937 bp wild-type bacteriophage T7 genome with 12,179 bp of engineered DNA using both synthetic DNA fragments and PCR-amplified T7 fragments, which contained all genetic elements of the 5′ end plus restriction enzyme sites.

Results

Conclusion