Programme(s) to which this project applies:
|☑ MPhil/PhD||☒ MRes[Med]||☒ URIS|
Protein engineering often requires changing a combination of amino acid sites chosen based on their structural information to significantly improve the activity of the protein. However, as the number of combinations increases exponentially with each amino acid site included, the protein engineering process can get very labor-intensive. Furthermore, the conventional selection process of protein variants, which requires sequencing each positively selected clone and comparing the selected protein to the wild-type protein, has very limited throughput, and, therefore, hinders us from repeating the selection process in different selection conditions. This leaves us not knowing whether we have selected the best variant in the most optimal selection condition.
Our lab developed the CombiSEAL screening platform (Choi et al., Nature Methods, 2019) to overcome the concerns mentioned above. The CombiSEAL platform modularizes a protein into multiple segments to generate mutant variants, where each of them is identified by a unique barcode. These variants are then seamlessly ligated into the protein-coding sequence while the corresponding barcodes are concatenated after the coding sequence. By using Next Generation Sequencing to sequence all these barcodes, we can get a quantitative readout for each variant; moreover, we can repeat the selection process of this barcoded library in multiple conditions until we find the most optimal condition.
The lab successfully identified multiple high-fidelity spCas9 variants by using the CombiSEAL platform (Choi et al., Nature Methods, 2019) and is interested in expanding the usage of the platform in engineering other gene-editing and therapeutic proteins.
This PhD project provides intensive training in high-throughput molecular biology, CRISPR-based genome editing, and NGS-based library screening and data analysis techniques.
Interested candidates should email Dr Alan Wong.
Dr ASL Wong, School of Biomedical Sciences
Dr Alan Siu-lun Wong is an Assistant Professor of the School of Biomedical Sciences at the University of Hong Kong (HKU). Before he joined HKU, he obtained his BSc and MPhil degrees in Biochemistry and Molecular Biotechnology from the Chinese University of Hong Kong in 2005 and 2007 respectively, and completed his PhD in Biochemistry at the Hong Kong University of Science and Technology in 2011. He joined the Synthetic Biology Group at Massachusetts Institute of Technology from 2012-2016 for postdoctoral training. Dr Wong was awarded with the Croucher Foundation Studentship (2008), the Butterfield-Croucher Award (2008), the Croucher Foundation Fellowship (2012), the Hong Kong Institution of Science Young Scientist Award in life science (2011), RGC Early Career Award (2016/2017), and NSFC Excellent Young Scientists Award (Hong Kong and Macau) (2020).
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