dc.contributor.author |
Singh, Advaita A
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dc.contributor.author |
Pillay, Priyen
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dc.contributor.author |
Naicker, Previn
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dc.contributor.author |
Alexandre, Kabamba B
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dc.contributor.author |
Malatji, Kanyane
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dc.contributor.author |
Mach, L
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dc.contributor.author |
Steinkellner, H
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dc.contributor.author |
Vorster, J
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dc.contributor.author |
Chikwamba, Rachel K
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dc.contributor.author |
Tsekoa, Tsepo L
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dc.date.accessioned |
2022-10-03T07:19:10Z |
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dc.date.available |
2022-10-03T07:19:10Z |
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dc.date.issued |
2022-08 |
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dc.identifier.citation |
Singh, A.A., Pillay, P., Naicker, P., Alexandre, K.B., Malatji, K., Mach, L., Steinkellner, H. & Vorster, J. et al. 2022. Transient proteolysis reduction of Nicotiana benthamiana-produced CAP256 broadly neutralizing antibodies using CRISPR/Cas9. <i>Frontiers in Plant Science, 13.</i> http://hdl.handle.net/10204/12501 |
en_ZA |
dc.identifier.uri |
DOI: 10.3389/fpls.2022.953654
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dc.identifier.uri |
http://hdl.handle.net/10204/12501
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dc.description.abstract |
The hypersensitive response is elicited by Agrobacterium infiltration of Nicotiana benthamiana, including the induction and accumulation of pathogenesis-related proteins, such as proteases. This includes the induction of the expression of several cysteine proteases from the C1 (papain-like cysteine protease) and C13 (legumain-like cysteine protease) families. This study demonstrates the role of cysteine proteases: NbVPE-1a, NbVPE-1b, and NbCysP6 in the proteolytic degradation of Nicotiana benthamiana (glycosylation mutant XTFT)-produced anti-human immunodeficiency virus broadly neutralizing antibody, CAP256-VRC26.25. Three putative cysteine protease cleavage sites were identified in the fragment crystallizable region. We further demonstrate the transient coexpression of CAP256-VRC26.25 with CRISPR/Cas9-mediated genome editing vectors targeting the NbVPE-1a, NbVPE-1b, and NbCysP6 genes which resulted in a decrease in CAP256-VRC26.25 degradation. No differences in structural features were observed between the human embryonic kidney 293 (HEK293)-produced and XTFT broadly neutralizing antibodies produced with and without the coexpression of genome-editing vectors. Furthermore, despite the presence of proteolytically degraded fragments of plant-produced CAP256-VRC26.25 without the coexpression of genome editing vectors, no influence on the in vitro functional activity was detected. Collectively, we demonstrate an innovative in planta strategy for improving the quality of the CAP256 antibodies through the transient expression of the CRISPR/Cas9 vectors. |
en_US |
dc.format |
Fulltext |
en_US |
dc.language.iso |
en |
en_US |
dc.relation.uri |
https://pubmed.ncbi.nlm.nih.gov/36061808/ |
en_US |
dc.source |
Frontiers in Plant Science, 13 |
en_US |
dc.subject |
Nicotiana benthamiana |
en_US |
dc.subject |
Genome editing |
en_US |
dc.subject |
Human immunodeficiency virus |
en_US |
dc.subject |
Immunoglobulin G |
en_US |
dc.subject |
Plant biotechnology |
en_US |
dc.subject |
Proteases |
en_US |
dc.title |
Transient proteolysis reduction of Nicotiana benthamiana-produced CAP256 broadly neutralizing antibodies using CRISPR/Cas9 |
en_US |
dc.type |
Article |
en_US |
dc.description.pages |
24 |
en_US |
dc.description.note |
© 2022 Singh, Pillay, Naicker, Alexandre, Malatji, Mach, Steinkellner, Vorster, Chikwamba and Tsekoa. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
en_US |
dc.description.cluster |
Chemicals |
en_US |
dc.description.cluster |
Next Generation Health |
en_US |
dc.description.cluster |
Leadership |
en_US |
dc.description.impactarea |
BT: Technology Demonstration |
en_US |
dc.description.impactarea |
Human Molecular Diagnostics |
en_US |
dc.description.impactarea |
Array Print Compan Diagnostics |
en_US |
dc.description.impactarea |
CSIR Executive Leadership Support |
en_US |
dc.description.impactarea |
BT: Technology Demonstration |
en_US |
dc.identifier.apacitation |
Singh, A. A., Pillay, P., Naicker, P., Alexandre, K. B., Malatji, K., Mach, L., ... Tsekoa, T. L. (2022). Transient proteolysis reduction of Nicotiana benthamiana-produced CAP256 broadly neutralizing antibodies using CRISPR/Cas9. <i>Frontiers in Plant Science, 13</i>, http://hdl.handle.net/10204/12501 |
en_ZA |
dc.identifier.chicagocitation |
Singh, Advaita A, Priyen Pillay, Previn Naicker, Kabamba B Alexandre, Kanyane Malatji, L Mach, H Steinkellner, J Vorster, Rachel K Chikwamba, and Tsepo L Tsekoa "Transient proteolysis reduction of Nicotiana benthamiana-produced CAP256 broadly neutralizing antibodies using CRISPR/Cas9." <i>Frontiers in Plant Science, 13</i> (2022) http://hdl.handle.net/10204/12501 |
en_ZA |
dc.identifier.vancouvercitation |
Singh AA, Pillay P, Naicker P, Alexandre KB, Malatji K, Mach L, et al. Transient proteolysis reduction of Nicotiana benthamiana-produced CAP256 broadly neutralizing antibodies using CRISPR/Cas9. Frontiers in Plant Science, 13. 2022; http://hdl.handle.net/10204/12501. |
en_ZA |
dc.identifier.ris |
TY - Article
AU - Singh, Advaita A
AU - Pillay, Priyen
AU - Naicker, Previn
AU - Alexandre, Kabamba B
AU - Malatji, Kanyane
AU - Mach, L
AU - Steinkellner, H
AU - Vorster, J
AU - Chikwamba, Rachel K
AU - Tsekoa, Tsepo L
AB - The hypersensitive response is elicited by Agrobacterium infiltration of Nicotiana benthamiana, including the induction and accumulation of pathogenesis-related proteins, such as proteases. This includes the induction of the expression of several cysteine proteases from the C1 (papain-like cysteine protease) and C13 (legumain-like cysteine protease) families. This study demonstrates the role of cysteine proteases: NbVPE-1a, NbVPE-1b, and NbCysP6 in the proteolytic degradation of Nicotiana benthamiana (glycosylation mutant XTFT)-produced anti-human immunodeficiency virus broadly neutralizing antibody, CAP256-VRC26.25. Three putative cysteine protease cleavage sites were identified in the fragment crystallizable region. We further demonstrate the transient coexpression of CAP256-VRC26.25 with CRISPR/Cas9-mediated genome editing vectors targeting the NbVPE-1a, NbVPE-1b, and NbCysP6 genes which resulted in a decrease in CAP256-VRC26.25 degradation. No differences in structural features were observed between the human embryonic kidney 293 (HEK293)-produced and XTFT broadly neutralizing antibodies produced with and without the coexpression of genome-editing vectors. Furthermore, despite the presence of proteolytically degraded fragments of plant-produced CAP256-VRC26.25 without the coexpression of genome editing vectors, no influence on the in vitro functional activity was detected. Collectively, we demonstrate an innovative in planta strategy for improving the quality of the CAP256 antibodies through the transient expression of the CRISPR/Cas9 vectors.
DA - 2022-08
DB - ResearchSpace
DP - CSIR
J1 - Frontiers in Plant Science, 13
KW - Nicotiana benthamiana
KW - Genome editing
KW - Human immunodeficiency virus
KW - Immunoglobulin G
KW - Plant biotechnology
KW - Proteases
LK - https://researchspace.csir.co.za
PY - 2022
T1 - Transient proteolysis reduction of Nicotiana benthamiana-produced CAP256 broadly neutralizing antibodies using CRISPR/Cas9
TI - Transient proteolysis reduction of Nicotiana benthamiana-produced CAP256 broadly neutralizing antibodies using CRISPR/Cas9
UR - http://hdl.handle.net/10204/12501
ER -
|
en_ZA |
dc.identifier.worklist |
25997 |
en_US |
dc.identifier.worklist |
CRISPR/Cas9 |
en_US |