Creation of a Bacterial Cell Controlled by a Chemically Synthesized Genome

Abstract

We report the design, synthesis, and assembly of the 1.08-mega-base pair Mycoplasma mycoides JCVI-syn1.0 genome starting from digitized genome sequence information and its transplantation into a M. capricolum recipient cell to create new M. mycoides cells that are controlled only by the synthetic chromosome. The only DNA in the cells is the designed synthetic DNA sequence, including "watermark" sequences and other designed gene deletions and polymorphisms, and mutations acquired during the building process. The new cells have expected phenotypic properties and are capable of continuous self-replication.

Building the blueprint of life

“…Efforts are under way in many labs throughout the world to produce a living strain with a minimal genome [1,7,8,14,[49][50][51]. These efforts are applying various approaches, depending on the organisms being used as a starting point and the specific scientific or industrial objectives motivating the effort.All the approaches can be classified as either top down or bottom up [4].…”

“…Top-down approaches involve starting with the genome of an existing (often far from minimal) organism and combining deletions to produce progressively smaller genomes [7,8,14,[49][50][51]. Bottom-up approaches involve starting with a very small genome and engineering a reduced version of the entire genome for implantation and viability testing [1]. An even more fundamental bottom-up approach is being pursued in which no natural genome is used as the starting point.…”

“…In the pure bottom-up approach, the minimal genome is designed computationally, synthesized in its entirety, and implanted in a living cell to produce a viable minimal organism. Every experimental step involved in implementing the bottom-up approach has now been successfully demonstrated with the genome of Mycoplasma mycoides as a template [1]. First, the M. mycoides genome was resequenced and computationally disassembled into 1078 overlapping cassettes, each 1080 BP long [1].…”

“…Every experimental step involved in implementing the bottom-up approach has now been successfully demonstrated with the genome of Mycoplasma mycoides as a template [1]. First, the M. mycoides genome was resequenced and computationally disassembled into 1078 overlapping cassettes, each 1080 BP long [1]. These cassettes were chemically synthesized and implanted in yeast, where the chromosome repair machinery of yeast was used to assemble these strands into a complete chromosome [9,52,53].…”

“…The recent breakthrough involving the de novo synthesis and implantation of an entire prokaryotic genome to create a living synthetic cell [1] represents a major triumph integrating decades of research in virtually every area of biology.This proofof-principle breakthrough has the potential to dramatically change how metabolic engineering is done. Rather than modifying existing organisms in small ways to fit them to our needs, entire genomes can be designed, synthesized, and implanted to produce living synthetic cells.…”