Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa
African swine fever virus (ASFV) is the etiological agent of a contagious and fatal disease of domestic pigs that has significant economic consequences for the global swine industry. Due to the lack of effective treatment and vaccines against African swine fever, there is an urgent need to leverage cutting-edge technologies and cost-effective approaches for generating and purifying recombinant virus to fast-track the development of live-attenuated ASFV vaccines. Here, we describe the use of the CRISPR/Cas9 gene editing and a cost-effective cloning system to produce recombinant ASFVs. Combining these approaches, we developed a recombinant virus lacking the non-essential gene A238L (5EL) in the highly virulent genotype IX ASFV (ASFV-Kenya-IX-1033) genome in less than 2 months as opposed to the standard homologous recombination with conventional purification techniques which takes up to 6 months on average. Our approach could therefore be a method of choice for less resourced laboratories in developing nations.
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Format: | Journal Article biblioteca |
Language: | English |
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Frontiers Media
2021-08-30
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Subjects: | african swine fever virus, animal diseases, genomics, crispr, |
Online Access: | https://hdl.handle.net/10568/114800 https://doi.org/10.3389/fgene.2021.733674 |
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dig-cgspace-10568-1148002023-12-08T19:36:04Z Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa Abkallo, Hussein M. Svitek, Nicholas Oduor, Bernard Awino, Elias Henson, Sonal P. Oyola, Samuel O. Mwalimu, Stephen Assad-Garcia, N. Fuchs, W. Vashee, S. Steinaa, Lucilla african swine fever virus animal diseases genomics crispr African swine fever virus (ASFV) is the etiological agent of a contagious and fatal disease of domestic pigs that has significant economic consequences for the global swine industry. Due to the lack of effective treatment and vaccines against African swine fever, there is an urgent need to leverage cutting-edge technologies and cost-effective approaches for generating and purifying recombinant virus to fast-track the development of live-attenuated ASFV vaccines. Here, we describe the use of the CRISPR/Cas9 gene editing and a cost-effective cloning system to produce recombinant ASFVs. Combining these approaches, we developed a recombinant virus lacking the non-essential gene A238L (5EL) in the highly virulent genotype IX ASFV (ASFV-Kenya-IX-1033) genome in less than 2 months as opposed to the standard homologous recombination with conventional purification techniques which takes up to 6 months on average. Our approach could therefore be a method of choice for less resourced laboratories in developing nations. 2021-08-30 2021-08-30T13:18:57Z 2021-08-30T13:18:57Z Journal Article Abkallo, H.M., Svitek, N., Oduor, B., Awino, E., Henson, S.P., Oyola, S.O., Mwalimu, S., Assad-Garcia, N., Fuchs, W., Vashee, S. and Steinaa, L. 2021. Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa. Frontiers in Genetics 12: 733674. 1664-8021 https://hdl.handle.net/10568/114800 https://doi.org/10.3389/fgene.2021.733674 en CC-BY-4.0 Open Access 733674 Frontiers Media Frontiers in Genetics |
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african swine fever virus animal diseases genomics crispr african swine fever virus animal diseases genomics crispr Abkallo, Hussein M. Svitek, Nicholas Oduor, Bernard Awino, Elias Henson, Sonal P. Oyola, Samuel O. Mwalimu, Stephen Assad-Garcia, N. Fuchs, W. Vashee, S. Steinaa, Lucilla Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa |
description |
African swine fever virus (ASFV) is the etiological agent of a contagious and fatal disease of domestic pigs that has significant economic consequences for the global swine industry. Due to the lack of effective treatment and vaccines against African swine fever, there is an urgent need to leverage cutting-edge technologies and cost-effective approaches for generating and purifying recombinant virus to fast-track the development of live-attenuated ASFV vaccines. Here, we describe the use of the CRISPR/Cas9 gene editing and a cost-effective cloning system to produce recombinant ASFVs. Combining these approaches, we developed a recombinant virus lacking the non-essential gene A238L (5EL) in the highly virulent genotype IX ASFV (ASFV-Kenya-IX-1033) genome in less than 2 months as opposed to the standard homologous recombination with conventional purification techniques which takes up to 6 months on average. Our approach could therefore be a method of choice for less resourced laboratories in developing nations. |
format |
Journal Article |
topic_facet |
african swine fever virus animal diseases genomics crispr |
author |
Abkallo, Hussein M. Svitek, Nicholas Oduor, Bernard Awino, Elias Henson, Sonal P. Oyola, Samuel O. Mwalimu, Stephen Assad-Garcia, N. Fuchs, W. Vashee, S. Steinaa, Lucilla |
author_facet |
Abkallo, Hussein M. Svitek, Nicholas Oduor, Bernard Awino, Elias Henson, Sonal P. Oyola, Samuel O. Mwalimu, Stephen Assad-Garcia, N. Fuchs, W. Vashee, S. Steinaa, Lucilla |
author_sort |
Abkallo, Hussein M. |
title |
Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa |
title_short |
Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa |
title_full |
Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa |
title_fullStr |
Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa |
title_full_unstemmed |
Rapid CRISPR/Cas9 editing of genotype IX African swine fever virus circulating in eastern and central Africa |
title_sort |
rapid crispr/cas9 editing of genotype ix african swine fever virus circulating in eastern and central africa |
publisher |
Frontiers Media |
publishDate |
2021-08-30 |
url |
https://hdl.handle.net/10568/114800 https://doi.org/10.3389/fgene.2021.733674 |
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