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Synthetic riboswitches promising tools for mammalian synthetic biology
RNA utilises many different mechanisms to control gene expression. Among the regulatory elements that respond to external stimuli, riboswitches are a prominent and elegant example. They are characteri
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Presentation
Synthetic riboswitches promising tools for mammalian synthetic biology00:00
RNA can adopt different conformations00:18
Riboswitches Use Alternative Structures00:47
Advantages of riboswitch regulation01:33
SELEX02:47
SELEX03:57
Synthetic RNA Biology in the Suess Lab04:29
Aptamer mediated translational inhibition05:20
Control of alternative splicing – Exon skipping06:49
Regulation of 5-Lipoxygenase08:26
Regulation of 5-Lipoxygenase09:03
Regulation of programmed cell death (apoptosis)09:19
Regulation of programmed cell death (apoptosis)09:56
Regulation of programmed cell death (apoptosis)10:35
Regulation of programmed cell death (apoptosis)10:50
CD20 controls the proper nanoscale receptor organization11:10
Tetracycline dependent ribozymes11:49
Tetracycline dependent ribozymes13:15
Tetracycline dependent ribozymes13:31
RNA synthetic biology13:38
What turns an aptamer into a riboswitch?14:39
What turns an aptamer into a riboswitch?14:50
A synthetic neomycin riboswitch15:19
Model for neomycin-induced conformational changes16:38
Ligand binding by conformational capture17:41
Ligand binding by conformational capture17:47
Ciprofloxacin binding aptamers18:27
Ciprofloxacin binding aptamers18:39
… and finally an aptamer with riboswitching activity 18:54
Improvement of candidate 10A18:59
What differs between an aptamer and a riboswitch?19:35
Stop flow measurements: dynamics of ligand binding19:47
Deep sequencing: slower enrichment of the riboswitch20:59
What turns an aptamer into an riboswitch21:48
Capture-SELEX22:39
The Tobramycin-Riboswitch23:43
The new one …. The Tobramycin-Riboswitch24:07
Is Capture-SELEX the Better Choice?24:57
Comparison: Capture vs Column-SELEX25:19
Doxycycline as new target …25:46
Doxycycline riboswitches25:46
Hybrid Riboswitches27:14
Hybrid Riboswitches Improvement of Dynamic Range27:54
Hybrid Riboswitches Implementation of a NAND gate28:22
Hybrid Riboswitches Implementation of a NAND gate28:54
NAND Gate Riboswitche29:44
NAND Gate Riboswitches30:49
Protein-Responsive Riboswitches31:17
Protein-Responsive Riboswitches31:56
Protein-Responsive Riboswitches32:28
Protein-Responsive Riboswitches32:47
An RNA as Alternative Activator of Gene Expression32:56
Protein – RNA Interaction33:45
Protein – RNA Interaction33:57
Control of Translation and Splicing34:05
Control of Translation and Splicing34:55
Control of Translation and Splicing35:05
The TetR Aptamer35:42
Control of 3’ splice site35:56
Control of nuclear import36:25
Control of nuclear import36:37
Control of nuclear import37:11
Bacterial repressors love RNA37:26
NagR and DasR binding aptamers37:50
NagR and DasR binding aptamers37:56
NagR and DasR binding aptamers38:08
Changing specificity from DasR towards NagR38:12
Changing specificity from DasR towards NagR38:45
Summary39:39
Danke41:07