Difference between revisions of "Team:MIT/Experiments/miRNA/more background"

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<h1 style="color:#000000; background-color:#FF9733; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center; font-family: Trebuchet MS"> Why use a miRNA sensor? </h1>
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The two most common methods for looking at the miRNA profile of a cell line are by miRNA microarray and quantitative polymerase chain reaction (qPCR). However, it has been indicated that the physical amount of miRNA present doesn't always have a strong correlation to the repression of the associated gene. By using a miRNA sensor, we can directly measure the activity of the miRNA, which the aspect of miRNA being used in the circuit.
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<i>"We detected the expression of more than 310 miRNAs (Fig. 2a). Our library included sensors for 165 of these miRNAs (188 when considering families), but we detected the suppression of only 67 sensors (Fig. 2b). Thus, 59% of the expressed miRNAs that we sampled did not have suppressive activity."</i> Mullokandov et. al Nature 2012
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<h1 style="color:#000000; background-color:#FF9733; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center; font-family: Trebuchet MS"> Using a miRNA Sensor </h1>
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The sensor, courtesy of Jeremy Gam, consists of a red fluorescent protein, mKate, which is constitutively controlled by the hef1a promoter. After mKate, you will notice here are four yellow blocks. These represent four miRNA target site domain repeats. After the target sites is a blue fluorescent protein also controlled by the hef1a promoter. When there is miRNA present, it will bind to the miRNA target site on the mKate mRNA and repress the expression of mKate.
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Revision as of 04:02, 14 October 2016

Description of Experiments

miRNA Candidates




Why use a miRNA sensor?


The two most common methods for looking at the miRNA profile of a cell line are by miRNA microarray and quantitative polymerase chain reaction (qPCR). However, it has been indicated that the physical amount of miRNA present doesn't always have a strong correlation to the repression of the associated gene. By using a miRNA sensor, we can directly measure the activity of the miRNA, which the aspect of miRNA being used in the circuit.

"We detected the expression of more than 310 miRNAs (Fig. 2a). Our library included sensors for 165 of these miRNAs (188 when considering families), but we detected the suppression of only 67 sensors (Fig. 2b). Thus, 59% of the expressed miRNAs that we sampled did not have suppressive activity." Mullokandov et. al Nature 2012







Using a miRNA Sensor


The sensor, courtesy of Jeremy Gam, consists of a red fluorescent protein, mKate, which is constitutively controlled by the hef1a promoter. After mKate, you will notice here are four yellow blocks. These represent four miRNA target site domain repeats. After the target sites is a blue fluorescent protein also controlled by the hef1a promoter. When there is miRNA present, it will bind to the miRNA target site on the mKate mRNA and repress the expression of mKate.