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Ribotale Essay

2818 words - 12 pages

Regulation of any biological phenotype is a complex and tuned phenomenon (Weng et al, 1999). Biological systems face frequent change in the environment, both at intrinsic and extrinsic levels. These systems have evolved themselves to integrate these fluctuations and take an appropriate decision (Helikar et al, 2008). Classical genetics and DNA sequencing have provided a lot of information about different types of genes and their functions, but how do these genes interact and integrate environmental signals to encode different phenotypic responses, is not clearly understood. Emergence of synthetic biology has enabled biologists to characterize these genetic components in isolation and predict the behavior of complex networks. Synthetic biology based approaches have been implemented to construct circuits mimicking natural circuits and understand the design principle of these circuits (Bashor et al, 2010; Rai et al, 2012). Synthetic gene networks have also been designed to perform several tasks including toggle switch, memory, oscillation, logic, biotechnological application (Danino et al, 2010; Gardner et al, 2000; Kobayashi et al, 2004; Siuti et al, 2013; Xie et al, 2011)
One of major hurdles in constructing complex synthetic gene circuits is availability of limited number of well characterized regulated promoters and cognate regulators. Most of the circuits have been constructed using PLac, PTet, PBAD and PcI promoters. Recently discovered, engineered CRISPR/Cas, TALE, and Riboswitches have been proved to be versatile tool for target specific gene regulation (Ceres et al, 2013; Mercer et al, 2013; Qi et al, 2013). CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated proteins) system is adaptive defense system against foreign nucleic acids present in several bacteria and archaea. Transcription activator-like effectors (TALEs) are secretory proteins synthesized by several species of Xanthomonas. TALEs regulate the gene expression in host plants involve in the colonization and survival of Xanthomonas. TALEs have a central highly conserved and tandem repeat domain. Each repeat comprised of 33 to 34 amino acids. Different repeats vary mostly at position 12 and 13. Amino acids present at these two positions are called repeat variable di-residues (RVDs). Each RVD has preference for a nucleotide. Repeats are modular and can be assembled in required way to bind with any target DNA sequence (Gaber et al, 2014; Meckler et al, 2013). TALEs are thus a powerful and modular tool, which can be engineered to target any DNA sequence. Riboswitches are non-protein coding regulatory RNA, present in 5´ untranslated (UTR) region of mRNA, that upon binding with small molecules or peptides undergo conformational changes to control gene expression at translational level. Riboswitches are conceptually comprised of two parts (i) ligand specific aptamer domain, and (ii) expression platform, which undergoes structural changes in response...

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