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The Aptamer Technology: Powerful Tools in Basic Science, Diagnostics and Therapy
Published on May 23, 20133790 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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Chapter list
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