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<p> The next step in developing our simulation was to calculate our protein concentration at any given time when using CRISPRi. Discussion with wet-lab revealed our method would be using CRISPRi as a repressor, which works by inhibiting the expression of one or more genes by binding to the promoter region <sup> 1 </sup>. The expanded mRNA and Protein concentration models from the Constitutive Gene Expression Model <sup> 2 </sup> were modified to include the element of repression from the CRISPRi inhibition. </p> | <p> The next step in developing our simulation was to calculate our protein concentration at any given time when using CRISPRi. Discussion with wet-lab revealed our method would be using CRISPRi as a repressor, which works by inhibiting the expression of one or more genes by binding to the promoter region <sup> 1 </sup>. The expanded mRNA and Protein concentration models from the Constitutive Gene Expression Model <sup> 2 </sup> were modified to include the element of repression from the CRISPRi inhibition. </p> | ||
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+ | <img src="https://static.igem.org/mediawiki/2017/e/e7/UNOT_crisprsystem.png";> | ||
<p> This system can be described as above. Where gRNA(i), Cas9, and mRNA are produced constitutively with their associated rates of production kc, kg, and k0 respectively. The Cas9 and gRNA(i) will undergo an inrreversible association to form Cas9:gRNA(i) at rate kf, which in turn inhibits the production of mRNA and reduce the production of Fluorescent protein (k1). All molecules spontaneously degrade and diffuse away at their own ssociated rate. (i) will account for us having multiple gRNAs and just as many fluorescent proteins i.e. i=3 with three fluorescent proteins and subsequent set of three gRNAs.</p> | <p> This system can be described as above. Where gRNA(i), Cas9, and mRNA are produced constitutively with their associated rates of production kc, kg, and k0 respectively. The Cas9 and gRNA(i) will undergo an inrreversible association to form Cas9:gRNA(i) at rate kf, which in turn inhibits the production of mRNA and reduce the production of Fluorescent protein (k1). All molecules spontaneously degrade and diffuse away at their own ssociated rate. (i) will account for us having multiple gRNAs and just as many fluorescent proteins i.e. i=3 with three fluorescent proteins and subsequent set of three gRNAs.</p> | ||
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Revision as of 01:45, 1 November 2017
MODELLING