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High content screening: from large libraries to functional hits
Published on Jun 28, 2019148 Views
Image-based high content screening is a powerful tool to gain systematic measurement of cell phenotype perturbation upon a given treatment. The combination of this technology with RNA interference, CR
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Chapter list
High content RNAi functional screening: from large libraries to functional hits00:00
How do you approach a biological problem?00:50
The photographer approach, The functional approach - 103:20
The photographer approach, The functional approach - 203:24
The photographer approach, The functional approach - 304:00
The functional approach 04:01
Screening Strategies, Requirements and tips - 104:39
Screening Strategies, Requirements and tips - 207:05
Screening Strategies, Requirements and tips - 307:06
High Content Screening facility at ICGEB Available libraries07:08
Automated liquid handling - 109:20
Automated liquid handling - 210:02
Molecular Devices ImageXpress Micro High-Content Screening Microscope10:04
PerkinElmer Operetta High-Content Screening Microscope10:31
High-Content Screening11:31
High-Throughput Screening11:32
Multidimensional Approach11:33
Data Analysis - 112:43
Data Analysis - 213:18
Data Analysis - 313:20
Data Analysis - 413:58
1. HTS-facility Active Projects14:12
2. HTS-facility Active Projects14:26
3. HTS-facility Active Projects14:29
Example 2_Multiwavelength Cell Scoring15:30
A high throughput screening assay to search for miRNAs enhancing HDR15:33
HTS for miRNAs enhancing HDR - 116:25
HTS for miRNAs enhancing HDR - 216:36
HTS for miRNAs enhancing HDR - 316:38
The selected miRNAs increase HDR in a cell line17:23
hsa-miR-4469 improves HDR efficiency in neonatal rat cardiomyocytes17:30
Example 3_Transfluor18:08
Disrimination19:12
Results - 119:49
Results - 220:06
Results - 320:17
Example 4_Multiwavelength Cell Translocation21:15
Adult cardiomyocytes do not proliferate22:52
Common degenerative conditions24:02
Time line showing the development of drugs against heart failure24:23
Biotherapeutics for myocardial ischemia and heart failure24:53
AAV vectors are outstanding tools for gene transfer24:56
Cardiac transduction after systemic injection of AAV9 vectors25:20
How to find novel biotherapeutics to regenerate the failing heart? - 125:21
How to find novel biotherapeutics to regenerate the failing heart? - 226:08
Modulate Sarcomere/Cytoplasm disasemmbly27:13
Screening for novel ubiquitin-conjugation factors - 127:38
Screening for novel ubiquitin-conjugation factors - 228:12
siRNAs regulating cardiomyocyte proliferation - 128:37
siRNAs regulating cardiomyocyte proliferation - 228:44
siRNAs regulating cardiomyocyte proliferation - 329:01
Silencing of top negative regulators also increases CMs cell number29:03
Adult heart functions upon depletion of Hit_1 - 129:40
Adult heart functions upon depletion of Hit_1 - 230:01
How to find novel biotherapeutics to regenerate the failing heart? - 338:09
Pathological cardiac remodeling38:30
Heart Failure - 138:32
Heart Failure - 338:34
Heart Failure - 238:54
Experimental Model39:59
The screening40:07
Hit Validation in Rat CMs40:29
Effects of in-vivo miR-665 over-expression under TAC - 141:32
Effects of in-vivo miR-665 over-expression under TAC - 242:30
Effects of in-vivo miR-665 over-expression under TAC - 342:38
Effects of in-vivo miR-665 over-expression under TAC - 442:56
How does it work?42:58
mRNA seq after cardiac AAV-hsa-miR665 over expression43:24
Functional siRNA screening44:36
FHL1, Enah and Xirp2 are direct targets of hsa-miR-66547:18
hsa-miR-665 over-expression decreases Young’s Modulus in primary NRVCMs48:29
Working Model49:31
Does hsa-miR-665 increase CM compliance to stretch?49:52
HFpEF - 149:56
HFpEF - 249:57
Working Model49:59
hsa-miR-665 levels associate with conserved EF in patients with dilated cardiomyopathy51:08
Thank you52:12