The Tools
The ambitious nature of projects in synthetic biology requires special methods to match them. For those projects that require extensive modification of genes and others perhaps whole genomes, conventional techniques used in molecular biology have to be im
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The Tools 5.1
Modularity and Standardization – 42
5.2
Reliability and Compatibility in Molecule Biology – 42
5.3
Establishing Standards – 43
5.4
The BioBrick Standard – 43
5.4.1 5.4.2 5.4.3
arts, Devices, and Systems – 45 P The BioBrick Foundation – 45 iGEM – 46
5.5
Discovery of the CRISPR/Cas Immune System – 46
5.6
dapting the CRISPR/Cas9 System for Molecular A Biology – 49
5.7
What Is It Capable Of? – 51
5.7.1 5.7.2 5.7.3
nock-In and Knock-Out Mutations – 51 K Gene Regulation – 52 Applying CRISPR/Cas9 to Genome Editing – 53
5.8
What Makes the CRISPR/Cas9 System Unique? – 53
5.9
What Dangers Does It Pose? – 53
5.10
How Are These Dangers Addressed? – 54
Further Reading – 56
© Springer International Publishing AG 2018 E.-K. Ehmoser-Sinner, C.-W. D. Tan, Lessons on Synthetic Bioarchitectures, Learning Materials in Biosciences, https://doi.org/10.1007/978-3-319-73123-0_5
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Chapter 5 · The Tools
What You Will Learn in This Chapter
5
The ambitious nature of projects in synthetic biology requires special methods to match them. For those projects that require extensive modification of genes and others perhaps whole genomes, conventional techniques used in molecular biology have to be improved to meet their needs. These needs include the necessity for gene modification methods to be reliable, easy to handle, and compatible between laboratories. This implies the need for certain standards, particularly for the materials and methods used. One approach to achieve this is to reduce the genetic material manipulated to highly interchangeable and interconnectable modules. We will look at how BioBricks allow us to do this. Another requirement is that the techniques used be precise and capable of large-scale changes to the target genetic material. We will see how the CRISPR/Cas9 system was developed to meet this need, as well as the range of DNA modifications it is capable of. Finally, the hazards posed by both technologies, as well as how those concerned have chosen to deal with them, are discussed.
5.1 Modularity and Standardization
As we have seen, synthetic biology hopes to play a significant role in the pharmaceutical industry. There are attempts to create novel antibiotics to tackle the challenge of rising resistance, new kinds of implants that would be more biocompatible, and new ways to target drugs more effectively, among other projects. However, a technology’s quality has to be taken to a higher level whenever it reaches industry. This is for reasons of safety, reliability and, very importantly, compatibility. When whole systems have to be redesigned to produce a new but related product, significant resources are wasted. Compatibility reduces this problem. Consider the Luer taper standard. As long as components contain the standard Luer male part, they can fit any other component that carries the Luer female part. This allows us to mix and match different components, easily giving us a wide range of solutions to a problem. Synthetic biology tries to
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