Circularly Permuted Far-Red Fluorescent Proteins

The color palette of genetically encoded fluorescent protein indicators (GEFPIs) has expanded rapidly in recent years. GEFPIs with excitation and emission within the “optical window” above 600 nm are expected to be superior in many aspects, such as enhanced tissue penetration, reduced autofluorescen...

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Autores principales:	Tianchen Wu, Yu Pang, Hui-wang Ai
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	far-red fluorescent protein circular permutation genetically encoded fluorescent biosensor pH sensitivity Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/728db4e6c848497c95ecc1a2274a4be1
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id	oai:doaj.org-article:728db4e6c848497c95ecc1a2274a4be1
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spelling	oai:doaj.org-article:728db4e6c848497c95ecc1a2274a4be12021-11-25T16:55:23ZCircularly Permuted Far-Red Fluorescent Proteins10.3390/bios111104382079-6374https://doaj.org/article/728db4e6c848497c95ecc1a2274a4be12021-11-01T00:00:00Zhttps://www.mdpi.com/2079-6374/11/11/438https://doaj.org/toc/2079-6374The color palette of genetically encoded fluorescent protein indicators (GEFPIs) has expanded rapidly in recent years. GEFPIs with excitation and emission within the “optical window” above 600 nm are expected to be superior in many aspects, such as enhanced tissue penetration, reduced autofluorescence and scattering, and lower phototoxicity. Circular permutation of fluorescent proteins (FPs) is often the first step in the process of developing single-FP-based GEFPIs. This study explored the tolerance of two far-red FPs, mMaroon1 and mCarmine, towards circular permutation. Several initial constructs were built according to previously reported circularly permuted topologies for other FP analogs. Mutagenesis was then performed on these constructs and screened for fluorescent variants. As a result, five circularly permuted far-red FPs (cpFrFPs) with excitation and emission maxima longer than 600 nm were identified. Some displayed appreciable brightness and efficient chromophore maturation. These cpFrFPs variants could be intriguing starting points to further engineer far-red GEFPIs for in vivo tissue imaging.Tianchen WuYu PangHui-wang AiMDPI AGarticlefar-red fluorescent proteincircular permutationgenetically encoded fluorescent biosensorpH sensitivityBiotechnologyTP248.13-248.65ENBiosensors, Vol 11, Iss 438, p 438 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	far-red fluorescent protein circular permutation genetically encoded fluorescent biosensor pH sensitivity Biotechnology TP248.13-248.65
spellingShingle	far-red fluorescent protein circular permutation genetically encoded fluorescent biosensor pH sensitivity Biotechnology TP248.13-248.65 Tianchen Wu Yu Pang Hui-wang Ai Circularly Permuted Far-Red Fluorescent Proteins
description	The color palette of genetically encoded fluorescent protein indicators (GEFPIs) has expanded rapidly in recent years. GEFPIs with excitation and emission within the “optical window” above 600 nm are expected to be superior in many aspects, such as enhanced tissue penetration, reduced autofluorescence and scattering, and lower phototoxicity. Circular permutation of fluorescent proteins (FPs) is often the first step in the process of developing single-FP-based GEFPIs. This study explored the tolerance of two far-red FPs, mMaroon1 and mCarmine, towards circular permutation. Several initial constructs were built according to previously reported circularly permuted topologies for other FP analogs. Mutagenesis was then performed on these constructs and screened for fluorescent variants. As a result, five circularly permuted far-red FPs (cpFrFPs) with excitation and emission maxima longer than 600 nm were identified. Some displayed appreciable brightness and efficient chromophore maturation. These cpFrFPs variants could be intriguing starting points to further engineer far-red GEFPIs for in vivo tissue imaging.
format	article
author	Tianchen Wu Yu Pang Hui-wang Ai
author_facet	Tianchen Wu Yu Pang Hui-wang Ai
author_sort	Tianchen Wu
title	Circularly Permuted Far-Red Fluorescent Proteins
title_short	Circularly Permuted Far-Red Fluorescent Proteins
title_full	Circularly Permuted Far-Red Fluorescent Proteins
title_fullStr	Circularly Permuted Far-Red Fluorescent Proteins
title_full_unstemmed	Circularly Permuted Far-Red Fluorescent Proteins
title_sort	circularly permuted far-red fluorescent proteins
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/728db4e6c848497c95ecc1a2274a4be1
work_keys_str_mv	AT tianchenwu circularlypermutedfarredfluorescentproteins AT yupang circularlypermutedfarredfluorescentproteins AT huiwangai circularlypermutedfarredfluorescentproteins
_version_	1718412843027529728

Circularly Permuted Far-Red Fluorescent Proteins

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