Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors

The spike (S) glycoprotein of SARS-Cov-2 facilitates viral entry into target cells via the cell surface receptor angiotensin-converting enzyme 2 (ACE2). Third generation HIV-1 lentiviral vectors can be pseudotyped to replace the native CD4 tropic envelope protein of the virus and thereby either limi...

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Autores principales:	Paul G. Ayoub, Arunima Purkayastha, Jason Quintos, Curtis Tam, Lindsay Lathrop, Kevin Tam, Marlene Ruiz, Roger P. Hollis, Brigitte N. Gomperts, Donald B. Kohn
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	SARS-CoV-2 pseudotype COVID-19 lentivirus air-liquid interface Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/efa1938c3f3c4807a4e0cd637ebbc8fb
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spelling	oai:doaj.org-article:efa1938c3f3c4807a4e0cd637ebbc8fb2021-12-01T06:59:34ZImproved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors2673-818X10.3389/fviro.2021.793320https://doaj.org/article/efa1938c3f3c4807a4e0cd637ebbc8fb2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fviro.2021.793320/fullhttps://doaj.org/toc/2673-818XThe spike (S) glycoprotein of SARS-Cov-2 facilitates viral entry into target cells via the cell surface receptor angiotensin-converting enzyme 2 (ACE2). Third generation HIV-1 lentiviral vectors can be pseudotyped to replace the native CD4 tropic envelope protein of the virus and thereby either limit or expand the target cell population. We generated a modified S glycoprotein of SARS-Cov-2 to pseudotype lentiviral vectors which efficiently transduced ACE2-expressing cells with high specificity and contain minimal off-target transduction of ACE2 negative cells. By utilizing optimized codons, modifying the S cytoplasmic tail domain, and including a mutant form of the spike protein, we generated an expression plasmid encoding an optimized protein that produces S-pseudotyped lentiviral vectors at an infectious titer (TU/mL) 1000-fold higher than the unmodified S protein and 4 to 10-fold more specific than the widely used delta-19 S-pseudotyped lentiviral vectors. S-pseudotyped replication-defective lentiviral vectors eliminate the need for biosafety-level-3 laboratories required when developing therapeutics against SARS-CoV-2 with live infectious virus. Furthermore, S-pseudotyped vectors with high activity and specificity may be used as tools to understand the development of immunity against SARS-CoV-2, to develop assays of neutralizing antibodies and other agents that block viral binding, and to allow in vivo imaging studies of ACE2-expressing cells.Paul G. AyoubArunima PurkayasthaJason QuintosCurtis TamLindsay LathropKevin TamMarlene RuizRoger P. HollisBrigitte N. GompertsBrigitte N. GompertsBrigitte N. GompertsDonald B. KohnDonald B. KohnDonald B. KohnDonald B. KohnDonald B. KohnFrontiers Media S.A.articleSARS-CoV-2pseudotypeCOVID-19lentivirusair-liquid interfaceMicrobiologyQR1-502ENFrontiers in Virology, Vol 1 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	SARS-CoV-2 pseudotype COVID-19 lentivirus air-liquid interface Microbiology QR1-502
spellingShingle	SARS-CoV-2 pseudotype COVID-19 lentivirus air-liquid interface Microbiology QR1-502 Paul G. Ayoub Arunima Purkayastha Jason Quintos Curtis Tam Lindsay Lathrop Kevin Tam Marlene Ruiz Roger P. Hollis Brigitte N. Gomperts Brigitte N. Gomperts Brigitte N. Gomperts Donald B. Kohn Donald B. Kohn Donald B. Kohn Donald B. Kohn Donald B. Kohn Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors
description	The spike (S) glycoprotein of SARS-Cov-2 facilitates viral entry into target cells via the cell surface receptor angiotensin-converting enzyme 2 (ACE2). Third generation HIV-1 lentiviral vectors can be pseudotyped to replace the native CD4 tropic envelope protein of the virus and thereby either limit or expand the target cell population. We generated a modified S glycoprotein of SARS-Cov-2 to pseudotype lentiviral vectors which efficiently transduced ACE2-expressing cells with high specificity and contain minimal off-target transduction of ACE2 negative cells. By utilizing optimized codons, modifying the S cytoplasmic tail domain, and including a mutant form of the spike protein, we generated an expression plasmid encoding an optimized protein that produces S-pseudotyped lentiviral vectors at an infectious titer (TU/mL) 1000-fold higher than the unmodified S protein and 4 to 10-fold more specific than the widely used delta-19 S-pseudotyped lentiviral vectors. S-pseudotyped replication-defective lentiviral vectors eliminate the need for biosafety-level-3 laboratories required when developing therapeutics against SARS-CoV-2 with live infectious virus. Furthermore, S-pseudotyped vectors with high activity and specificity may be used as tools to understand the development of immunity against SARS-CoV-2, to develop assays of neutralizing antibodies and other agents that block viral binding, and to allow in vivo imaging studies of ACE2-expressing cells.
format	article
author	Paul G. Ayoub Arunima Purkayastha Jason Quintos Curtis Tam Lindsay Lathrop Kevin Tam Marlene Ruiz Roger P. Hollis Brigitte N. Gomperts Brigitte N. Gomperts Brigitte N. Gomperts Donald B. Kohn Donald B. Kohn Donald B. Kohn Donald B. Kohn Donald B. Kohn
author_facet	Paul G. Ayoub Arunima Purkayastha Jason Quintos Curtis Tam Lindsay Lathrop Kevin Tam Marlene Ruiz Roger P. Hollis Brigitte N. Gomperts Brigitte N. Gomperts Brigitte N. Gomperts Donald B. Kohn Donald B. Kohn Donald B. Kohn Donald B. Kohn Donald B. Kohn
author_sort	Paul G. Ayoub
title	Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors
title_short	Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors
title_full	Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors
title_fullStr	Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors
title_full_unstemmed	Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors
title_sort	improved sars-cov-2 spike glycoproteins for pseudotyping lentiviral vectors
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/efa1938c3f3c4807a4e0cd637ebbc8fb
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Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors

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