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The Aptamer Technology: Powerful Tools in Basic Science, Diagnostics and Therapy
Published on 2013-05-233805 Views
DNA and RNA aptamers interact with their target proteins with high selectivity, specificity and affinity. The SELEX method consists of iterative cycles of in vitro screening of a combinatorial oligonu
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Presentation
The Aptamer Technology: Powerful Tools in Basic Science, Diagnostics and Therapy00:00
SELEX00:32
What is an Aptamer?01:16
Aptamers02:18
Isolation of Short Oligonucleotides (Aptamers)04:44
Aptamer Development06:31
Direct / Indirect Recognition08:22
In VitroSelection of High-affinity RNA Ligands09:20
The Original Experiment: Selection of RNA Molecules that Bind to T4 DNA-polymerase10:53
Natural xIn vitroEvolution13:02
Why to Use Nucleic Acids?14:52
Chemical Modification of the Ribose of Pyrimidines Results in Nuclease-resistant Aptamers16:23
Aptamers are easily developed for transcription factors and other soluble proteins17:41
Example 1:Aptamers Binding to the Cocaine-site of the Nicotinic Acetylcholine Receptor19:08
Abused Drugs, Toxins and Compounds of Therapeutic Importance that Inhibit the Nicotinic Acetylcholine receptor (nAChR)20:15
Structure of the Nicotinic Acetylcholine Receptor (nAChR)21:21
SELEX for Aptamers Binding to Membrane Proteins21:56
Gel-Shift Comparison of SELEX Pools23:33
Reiterative SELEX Cycles Result in Nanomolar Binding Affinity of selected RNA molecules24:14
Aptamers Containing Consensus Sequences24:47
Displacement of Class I and II Aptamers by Cocaine25:20
Part 1: Proof of Priniciple26:28
The Protector26:59
Example 2: Development of Aptamers as Blockers of Host Cell Invasion by Trypanosoma cruzi29:34
Chagas’ Disease in Brazil and Latin America29:58
Receptor-ligand Interactions30:53
Host Cell-matrix Molecules Bind with High Affinity to their Receptors on T. cruzi Trypomastigotes32:02
SELEX Procedure for the Evolution of RNA Aptamers32:46
The Selected RNA Molecules Bind Mainly to their Targets34:36
Displacement of the SELEX 7 RNA Pool36:17
Consensus Sequences of Aptamers Binding to T. cruzisurfaces36:57
Saturation Analysis of Consensus Aptamer Binding37:30
Will Aptamers Interfere with Cell Infection?37:48
The selected RNA Aptamers Partially Inhibit Invasion39:15
Actual projects40:07
Aptamers as Specific Inhibitors of P2Y2 Receptors41:41
Purinergic P2X and P2Y Receptor Activation41:59
P2Y2 receptors42:49
Aptamer Selection for P2Y2 Receptors43:26
Identified anti-P2Y2Aptamers have Conserved Sequence Motifs44:39
Selected aptamer B7 Preferentially Binds to P2Y2 Receptors45:15
Aptamers Abolish P2Y2R-mediated Protection of Embryonal Carcinoma Cells Against Apoptosis46:45
Aptamers for Isolation of Adipose-derived Mesenchymal Stem Cells48:47
Isolation of Stem Cells Based on Marker Expression Profiles49:29
Aptamer SELEX for human stem cell specific ligands50:53
Three initial cycles of stem cell SELEX52:31
Aptamers for Recognition of Mesenchymal Stem Cells in Adipose Tissue53:44
Aptamers can Substitute CD29+/CD90+MSC Labeling54:06
Aptamers Purify Human Adipose Stem Cells56:22
Differentiation of Purified Adipose-MSCs into Neural Phenotypes57:38
Aptamers Label subpopulations within Stem Cells58:52
Cloned Aptamers are Screened for Novel Specific Markers of Mesenchymal Stem Cells59:33
Aptamers Label Plasmodium falciparum-infected Erythrocytes in Live Cell Imaging01:00:49
In vitro Evolution of RNA Molecules for Therapeutic Applications01:03:47
Development into Therapeutics01:06:00
Development of an anti-VEGF Aptamer01:08:11
Aptamers vs. other therapeutics01:09:05
Acknowledgments01:09:46