Publication:
Identification of modulators of chemotherapeutic resistance using a random shRNA library

dc.contributor.advisor Arndt, Greg en_US
dc.contributor.advisor Dawes, Ian en_US
dc.contributor.author Ramadas, Radhika en_US
dc.date.accessioned 2022-03-21T11:48:08Z
dc.date.available 2022-03-21T11:48:08Z
dc.date.issued 2012 en_US
dc.description.abstract High-throughput gene silencing using RNA interference (RNAi) libraries is a rapidly expanding strategy to study the molecular pathways underlying human diseases. A majority of RNAi libraries contain expression constructs designed to target specific genes or gene sub-classes. The use of random RNAi libraries offers a more wide-reaching alternative to identify novel therapeutic targets. However, most of the previously reported protocols for the generation of random RNAi libraries remain highly complex. In this thesis, the development of a simplified and efficient method for the construction of a random shRNA-encoding library is reported. To prove the functionality of the random shRNA-encoding library, it was used to investigate the molecular components and mechanisms used by cancer cells to acquire resistance to three anti-cancer drugs from two drug classes with distinct mechanisms of action. Using independent selection assays, cells transduced with the random shRNA-encoding library were exposed to cisplatin, carboplatin or paclitaxel, and resistant cells isolated. A total of 159 unique shRNA-encoding inserts were recovered, sequenced and used to perform homology-based searching of protein- and RNA-encoding databases. Several of the inserts were enriched in cells displaying resistance to the specific chemotherapeutic agents. The predicted target sequences included both previously reported targets implicated in cellular resistance mechanisms and novel targets, thus proving the feasibility of this approach in identifying new targets for therapeutic agents. The cell-based RNAi screen and subsequent bioinformatic analysis produced a list of novel gene targets for further investigation of the role each plays in chemotherapeutic drug resistance. en_US
dc.identifier.uri http://hdl.handle.net/1959.4/52276
dc.language English
dc.language.iso EN en_US
dc.publisher UNSW, Sydney en_US
dc.rights CC BY-NC-ND 3.0 en_US
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/3.0/au/ en_US
dc.subject.other Chemotherapeutic drug resistance en_US
dc.subject.other RNA interference en_US
dc.subject.other Random shRNA-encoding library en_US
dc.title Identification of modulators of chemotherapeutic resistance using a random shRNA library en_US
dc.type Thesis en_US
dcterms.accessRights open access
dcterms.rightsHolder Ramadas, Radhika
dspace.entity.type Publication en_US
unsw.accessRights.uri https://purl.org/coar/access_right/c_abf2
unsw.identifier.doi https://doi.org/10.26190/unsworks/15837
unsw.relation.faculty Science
unsw.relation.originalPublicationAffiliation Ramadas, Radhika, Biotechnology & Biomolecular Sciences, Faculty of Science, UNSW en_US
unsw.relation.originalPublicationAffiliation Arndt, Greg, Children's Cancer Institute Australia for Medical Research, UNSW en_US
unsw.relation.originalPublicationAffiliation Dawes, Ian, Biotechnology & Biomolecular Sciences, Faculty of Science, UNSW en_US
unsw.relation.school School of Biotechnology & Biomolecular Sciences *
unsw.thesis.degreetype PhD Doctorate en_US
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