About the Project
The Department of Life Sciences at Imperial College London has funding for 7 3-year departmentally funded PhD studentships, to commence October 2021. Please see below for the details of the individual projects, 6 of which involve interactions with industry. For information on the individual projects please contact the relevant supervisor as detailed below. For queries about the application and interview process please contact Rozan Hamilton-Nixon at [Email Address Removed].
Application deadline: 12noon, 5 April 2021
Application process: To apply please complete the application form found at https://www.imperial.ac.uk/bbsrc-doctoral-training-partnership/application-process/application-form/[GK1]
Dr. Tony Southall. [Email Address Removed]
The ability to modify and tag endogenously expressed genes is an essential tool in many areas of academic research and the biotechnology industry. The common and often used approach, CRISPR/Cas mediated homologous recombination, has many drawbacks which include off-target effects, unpredictable locus specific on-target efficiency, potentially unwanted introduction of DNA concatemers, requires laborious clonal isolation followed by thorough genotypic analysis and delivery of CRISPR/Cas reagents. Here, we propose to further develop and optimise an alternative tool that instead reprograms the mRNA, rather than editing the DNA. This method is based on the expression of a pre-trans-splicing molecule (PTM), which is an RNA that binds to the intron of a pre-mRNA (before splicing), leading to trans-splicing to the PTM, which contains the reprogrammed exons. Previous studies, including our own, have validated this method works. However, its general applicability is limited by low efficiency, off-target effects, and leaky translation of the PTM. Here we will test and optimise new modifications that can address these issues. Initial modifications and novel applications will be optimised and tested in Drosophila, allowing for definitive testing for any off-target reprogramming events. These technologies will then be optimised for use in identifying programmable differentiated cell populations from human iPSC cells through our partnership with bit.bio (The Cell Coding Company), Cambridge, UK.
2. Trochet D, Prudhon B, Jollet A, Lorain S, Bitoun M. Reprogramming the Dynamin 2 mRNA by Spliceosome-mediated RNA Trans-splicing. Mol Ther Nucleic Acids. 2016 Sep 13;5(9):e362. doi: 10.1038/mtna.2016.67. PMID: 27623444; PMCID: PMC5056991.
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