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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Main Authors: 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
Format: Journal Article biblioteca
Language:English
Published: Frontiers Media 2021-08-30
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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spelling 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
institution CGIAR
collection DSpace
country Francia
countrycode FR
component Bibliográfico
access En linea
databasecode dig-cgspace
tag biblioteca
region Europa del Oeste
libraryname Biblioteca del CGIAR
language English
topic african swine fever virus
animal diseases
genomics
crispr
african swine fever virus
animal diseases
genomics
crispr
spellingShingle 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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