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| Status |
Public on Apr 09, 2019 |
| Title |
Estimating Genome-wide Off-Target Effects for Pyrrole-Imidazole Polyamide Binding by a Pathway-Based Expression Profiling Approach (SiHA cells) |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Side and off-target effects remain a difficult issue in the search of pharmaceutical leads, especially with DNA-binding molecules or genome editing methods where the issue becomes particularly thorny. A particular case is the investigations into the off-target effects of N-methylpyrrole-N-methylimidazole polyamides, a naturally inspired class of DNA binders that discriminately access the minor groove with strong affinity and sequence specificity but the relatively short motif of < 20 bases often imply possible non-unique genomic binding. Multiple binding sites may translate to binding at non-intended loci, potentially leading to off-target effects, issues that very few approaches are able to address to-date. We here report an analytical method of inferring off-target binding from expression profiling based on the relative impact to various biochemical pathways, as well as an accompanying side effect prediction engine to allow candidate polyamides to be systematically screened. This method marks the first attempt in PI polyamide research to identify elements in various biochemical pathways sensitive to the treatment of a candidate polyamide to infer possible off-target effects. Expression changes were then considered for their possible effect on outward phenotypic changes, manifested as side effects, should the same PI polyamide candidate be administered clinically. We also performed a series of animal experiments to validate some of these effects, and found some corroboration in certain side effect manifestations, such as changes in the level of aspartate transaminase in ICR and nude mice after injection of some of the candidate polyamides in this study.
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| Overall design |
O(4) to O(5) cells per condition were plated in a 6-well microtiter plate for overnight attachment prior to treatment of candidate polyamides or 0.05% DMSO. After RNA extraction with RNeasy Plus Mini Kit (Qiagen), labelled with RNA Spike-In Kit and analyzed on SurePrint G3 Human GE 8x60K V2 microarrays.
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| Contributor(s) |
Lin J, Krishnamurthy S, Atsushi T |
| Citation(s) |
30964912 |
| Submission date |
Dec 28, 2018 |
| Last update date |
Apr 14, 2019 |
| Contact name |
Jason Lin |
| Organization name |
Chiba Cancer Center Research Institute
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| Street address |
666-2 Nitona-cho, Chuo-ku
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| City |
Chiba |
| State/province |
Chiba |
| ZIP/Postal code |
260-8717 |
| Country |
Japan |
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| Platforms (1) |
| GPL16699 |
Agilent-039494 SurePrint G3 Human GE v2 8x60K Microarray 039381 (Feature Number version) |
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| Samples (5)
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| This SubSeries is part of SuperSeries: |
| GSE124462 |
Estimating Genome-wide Off-Target Effects for Pyrrole-Imidazole Polyamide Binding by a Pathway-Based Expression Profiling Approach |
|
| Relations |
| BioProject |
PRJNA512086 |
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