Abstract
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.
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Further Reading
Baltimore D, Berg P, Botchan M, Carroll D, Charo RA, Church G, Corn JE, Daley GQ, Doudna JA, Fenner M et al (2015) Biotechnology. A prudent path forward for genomic engineering and germline gene modification. Science 348:36–38
Casini A, Storch M, Baldwin GS, Ellis T (2015) Bricks and blueprints: methods and standards for DNA assembly. Nat Rev Mol Cell Biol 16:568–576
Doudna J (2015) Genome-editing revolution: my whirlwind year with CRISPR. Nature 528:469–471
Doudna JA, Charpentier E (2014) Genome editing. The new frontier of genome engineering with CRISPR-Cas9. Science 346:1258096
Freedman BS, Brooks CR, Lam AQ, Fu H, Morizane R, Agrawal V, Saad AF, Li MK, Hughes MR, Werff RV et al (2015) Modelling kidney disease with CRISPR-mutant kidney organoids derived from human pluripotent epiblast spheroids. Nat Commun 6:8715
Hart T, Chandrashekhar M, Aregger M, Steinhart Z, Brown KR, MacLeod G, Mis M, Zimmermann M, Fradet-Turcotte A, Sun S et al (2015) High-resolution CRISPR screens reveal fitness genes and genotype-specific cancer liabilities. Cell 163:1515–1526
Ho-Shing O, Lau KH, Vernon W, Eckdahl TT, Campbell AM (2012) Assembly of standardized DNA parts using BioBrick ends in E. coli. Methods Mol Biol 852:61–76
Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337:816–821
Lanphier E, Urnov F, Haecker SE, Werner M, Smolenski J (2015) Don’t edit the human germ line. Nature 519:410–411
Liang P, Xu Y, Zhang X, Ding C, Huang R, Zhang Z, Lv J, Xie X, Chen Y, Li Y et al (2015) CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes. Protein Cell 6:363–372
Merkle FT, Neuhausser WM, Santos D, Valen E, Gagnon JA, Maas K, Sandoe J, Schier AF, Eggan K (2015) Efficient CRISPR-Cas9-mediated generation of knockin human pluripotent stem cells lacking undesired mutations at the targeted locus. Cell Rep 11:875–883
Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG et al (2014) Genome-scale CRISPR-Cas9 knockout screening in human cells. Science 343:84–87
Shetty RP, Endy D, Knight TF Jr (2008) Engineering BioBrick vectors from BioBrick parts. J Biol Eng 2:5
Wang H, Yang H, Shivalila CS, Dawlaty MM, Cheng AW, Zhang F, Jaenisch R (2013) One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153:910–918
Yang L, Guell M, Niu D, George H, Lesha E, Grishin D, Aach J, Shrock E, Xu W, Poci J et al (2015) Genome-wide inactivation of porcine endogenous retroviruses (PERVs). Science 350:1101–1104
Zhu W, Lei R, Le Duff Y, Li J, Guo F, Wainberg MA, Liang C (2015) The CRISPR/Cas9 system inactivates latent HIV-1 proviral DNA. Retrovirology 12:22
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Ehmoser-Sinner, EK., Tan, CW.D. (2018). The Tools. In: Lessons on Synthetic Bioarchitectures. Learning Materials in Biosciences. Springer, Cham. https://doi.org/10.1007/978-3-319-73123-0_5
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DOI: https://doi.org/10.1007/978-3-319-73123-0_5
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