Emerging tools for synthetic genome design

Abstract

Synthetic biology is an emerging discipline for designing and synthesizing predictable, measurable, controllable, and transformable biological systems. These newly designed biological systems have great potential for the development of cheaper drugs, green fuels, biodegradable plastics, and targeted cancer therapies over the coming years. Fortunately, our ability to quickly and accurately engineer biological systems that behave predictably has been dramatically expanded by significant advances in DNA-sequencing, DNA-synthesis, and DNA-editing technologies. Here, we review emerging technologies and methodologies in the field of building designed biological systems, and we discuss their future perspectives.

Fig. 1.

Overall scheme and related synthetic biology tools for building new biological systems.

Fig. 2.

Genome assembly methods. (A) Standard assembly with restriction enzymes, (B) In-fusion assembly, (C) Sequence and ligase independent cloning (SLIC), (D) Gibson assembly, and (E) Golden Gate assembly.

Fig. 3.

Schematic diagram of genome editing methods. (A) Zinc-finger nuclease (ZFNs), which is recognized and bound with three nucleotides of the genome; (B) Transcription activator-like effector nuclease (TALENs), in which each amino acid in the DNA-binding domain recognizes one nucleotide; and (C) Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system, which uses guide RNAs for recognizing target DNA.