Development and application of genetic engineering tools

SNPs in the genomic DNA are linked to numerous diseases. In these cases, the disease and healthy alleles differ by a single DNA base. Using CRISPR-Cas9 base editors, it is possible to modify SNPs by converting the targeted disease-related DNA base into another base. CGBEs are different from CBEs and...

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Main Author: Teo, Yi Yong
Other Authors: Tan Meng How
Format: Final Year Project
Language:English
Published: Nanyang Technological University 2022
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Online Access:https://hdl.handle.net/10356/159073
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-1590732022-06-09T23:58:50Z Development and application of genetic engineering tools Teo, Yi Yong Tan Meng How School of Chemical and Biomedical Engineering mh.tan@ntu.edu.sg Engineering::Bioengineering SNPs in the genomic DNA are linked to numerous diseases. In these cases, the disease and healthy alleles differ by a single DNA base. Using CRISPR-Cas9 base editors, it is possible to modify SNPs by converting the targeted disease-related DNA base into another base. CGBEs are different from CBEs and ABEs, and can be used to convert C:G base pairs to G:C base pairs. CGBEs can be packaged into AAV vectors as a means of delivery to reach the target cells in vivo. AAV vectors are used due to its low pathogenic properties. However, AAV vectors have a packaging limit of ~5kb. This means that the CGBE to be packaged into the AAV vector needs to be below this packaging limit to have the intended function. Thus, there is a need to truncate the CGBE such that it can be packaged into the AAV vector. However, truncating the CGBE would mean that segments of DNA in CGBE would have to be removed. Therefore, to ensure that the CGBE construct would continue to function as intended after the removal of the DNA segments, the editing efficiencies of the truncated CGBE are investigated. Different deletions of DNA segments at various positions of the CGBE construct are also investigated to generate a truncated CGBE with relatively high editing efficiency. The experiment determined the combinations of deletions in relation to the editing efficiency of the contructs. Bachelor of Engineering (Bioengineering) 2022-06-09T06:29:29Z 2022-06-09T06:29:29Z 2022 Final Year Project (FYP) Teo, Y. Y. (2022). Development and application of genetic engineering tools. Final Year Project (FYP), Nanyang Technological University, Singapore. https://hdl.handle.net/10356/159073 https://hdl.handle.net/10356/159073 en application/pdf Nanyang Technological University
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Engineering::Bioengineering
spellingShingle Engineering::Bioengineering
Teo, Yi Yong
Development and application of genetic engineering tools
description SNPs in the genomic DNA are linked to numerous diseases. In these cases, the disease and healthy alleles differ by a single DNA base. Using CRISPR-Cas9 base editors, it is possible to modify SNPs by converting the targeted disease-related DNA base into another base. CGBEs are different from CBEs and ABEs, and can be used to convert C:G base pairs to G:C base pairs. CGBEs can be packaged into AAV vectors as a means of delivery to reach the target cells in vivo. AAV vectors are used due to its low pathogenic properties. However, AAV vectors have a packaging limit of ~5kb. This means that the CGBE to be packaged into the AAV vector needs to be below this packaging limit to have the intended function. Thus, there is a need to truncate the CGBE such that it can be packaged into the AAV vector. However, truncating the CGBE would mean that segments of DNA in CGBE would have to be removed. Therefore, to ensure that the CGBE construct would continue to function as intended after the removal of the DNA segments, the editing efficiencies of the truncated CGBE are investigated. Different deletions of DNA segments at various positions of the CGBE construct are also investigated to generate a truncated CGBE with relatively high editing efficiency. The experiment determined the combinations of deletions in relation to the editing efficiency of the contructs.
author2 Tan Meng How
author_facet Tan Meng How
Teo, Yi Yong
format Final Year Project
author Teo, Yi Yong
author_sort Teo, Yi Yong
title Development and application of genetic engineering tools
title_short Development and application of genetic engineering tools
title_full Development and application of genetic engineering tools
title_fullStr Development and application of genetic engineering tools
title_full_unstemmed Development and application of genetic engineering tools
title_sort development and application of genetic engineering tools
publisher Nanyang Technological University
publishDate 2022
url https://hdl.handle.net/10356/159073
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