High‐throughput, microscopy‐based screening and quantification of genetic elements

Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screeni...
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Autor*in:	Rongrong Zhang [verfasserIn] Yajia Huang [verfasserIn] Mei Li [verfasserIn] Lei Wang [verfasserIn] Bing Li [verfasserIn] Aiguo Xia [verfasserIn] Ye Li [verfasserIn] Shuai Yang [verfasserIn] Fan Jin [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2023

Schlagwörter:	characterization methods high‐throughput robust genetic elements synthetic biology

Übergeordnetes Werk:	In: mLife - Wiley, 2022, 2(2023), 4, Seite 450-461
Übergeordnetes Werk:	volume:2 ; year:2023 ; number:4 ; pages:450-461

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1002/mlf2.12096

Katalog-ID:	DOAJ097981982

Internformat


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520			\|a Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs.
650		4	\|a characterization methods
650		4	\|a high‐throughput
650		4	\|a robust genetic elements
650		4	\|a synthetic biology
653		0	\|a Microbiology
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700	0		\|a Mei Li \|e verfasserin \|4 aut
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700	0		\|a Bing Li \|e verfasserin \|4 aut
700	0		\|a Aiguo Xia \|e verfasserin \|4 aut
700	0		\|a Ye Li \|e verfasserin \|4 aut
700	0		\|a Shuai Yang \|e verfasserin \|4 aut
700	0		\|a Fan Jin \|e verfasserin \|4 aut
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912			\|a GBV_ILN_40
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912			\|a GBV_ILN_65
912			\|a GBV_ILN_69
912			\|a GBV_ILN_70
912			\|a GBV_ILN_73
912			\|a GBV_ILN_74
912			\|a GBV_ILN_95
912			\|a GBV_ILN_105
912			\|a GBV_ILN_110
912			\|a GBV_ILN_151
912			\|a GBV_ILN_161
912			\|a GBV_ILN_170
912			\|a GBV_ILN_171
912			\|a GBV_ILN_213
912			\|a GBV_ILN_224
912			\|a GBV_ILN_230
912			\|a GBV_ILN_285
912			\|a GBV_ILN_293
912			\|a GBV_ILN_602
912			\|a GBV_ILN_636
912			\|a GBV_ILN_2004
912			\|a GBV_ILN_2005
912			\|a GBV_ILN_2006
912			\|a GBV_ILN_2007
912			\|a GBV_ILN_2010
912			\|a GBV_ILN_2011
912			\|a GBV_ILN_2014
912			\|a GBV_ILN_2026
912			\|a GBV_ILN_2027
912			\|a GBV_ILN_2034
912			\|a GBV_ILN_2037
912			\|a GBV_ILN_2038
912			\|a GBV_ILN_2044
912			\|a GBV_ILN_2048
912			\|a GBV_ILN_2049
912			\|a GBV_ILN_2050
912			\|a GBV_ILN_2055
912			\|a GBV_ILN_2056
912			\|a GBV_ILN_2057
912			\|a GBV_ILN_2059
912			\|a GBV_ILN_2061
912			\|a GBV_ILN_2064
912			\|a GBV_ILN_2068
912			\|a GBV_ILN_2088
912			\|a GBV_ILN_2106
912			\|a GBV_ILN_2108
912			\|a GBV_ILN_2110
912			\|a GBV_ILN_2111
912			\|a GBV_ILN_2118
912			\|a GBV_ILN_2122
912			\|a GBV_ILN_2143
912			\|a GBV_ILN_2144
912			\|a GBV_ILN_2147
912			\|a GBV_ILN_2148
912			\|a GBV_ILN_2152
912			\|a GBV_ILN_2153
912			\|a GBV_ILN_2232
912			\|a GBV_ILN_2336
912			\|a GBV_ILN_2470
912			\|a GBV_ILN_2507
912			\|a GBV_ILN_2522
912			\|a GBV_ILN_4012
912			\|a GBV_ILN_4035
912			\|a GBV_ILN_4037
912			\|a GBV_ILN_4046
912			\|a GBV_ILN_4112
912			\|a GBV_ILN_4125
912			\|a GBV_ILN_4126
912			\|a GBV_ILN_4242
912			\|a GBV_ILN_4249
912			\|a GBV_ILN_4251
912			\|a GBV_ILN_4305
912			\|a GBV_ILN_4306
912			\|a GBV_ILN_4307
912			\|a GBV_ILN_4313
912			\|a GBV_ILN_4322
912			\|a GBV_ILN_4323
912			\|a GBV_ILN_4324
912			\|a GBV_ILN_4325
912			\|a GBV_ILN_4326
912			\|a GBV_ILN_4333
912			\|a GBV_ILN_4334
912			\|a GBV_ILN_4335
912			\|a GBV_ILN_4336
912			\|a GBV_ILN_4338
912			\|a GBV_ILN_4367
912			\|a GBV_ILN_4700
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allfields	10.1002/mlf2.12096 doi (DE-627)DOAJ097981982 (DE-599)DOAJ21f12515f24747119dd1548341552373 DE-627 ger DE-627 rakwb eng QR1-502 Rongrong Zhang verfasserin aut High‐throughput, microscopy‐based screening and quantification of genetic elements 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs. characterization methods high‐throughput robust genetic elements synthetic biology Microbiology Yajia Huang verfasserin aut Mei Li verfasserin aut Lei Wang verfasserin aut Bing Li verfasserin aut Aiguo Xia verfasserin aut Ye Li verfasserin aut Shuai Yang verfasserin aut Fan Jin verfasserin aut In mLife Wiley, 2022 2(2023), 4, Seite 450-461 (DE-627)1804448230 (DE-600)3120417-X 2770100X nnns volume:2 year:2023 number:4 pages:450-461 https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/article/21f12515f24747119dd1548341552373 kostenfrei https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/toc/2770-100X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2023 4 450-461
spelling	10.1002/mlf2.12096 doi (DE-627)DOAJ097981982 (DE-599)DOAJ21f12515f24747119dd1548341552373 DE-627 ger DE-627 rakwb eng QR1-502 Rongrong Zhang verfasserin aut High‐throughput, microscopy‐based screening and quantification of genetic elements 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs. characterization methods high‐throughput robust genetic elements synthetic biology Microbiology Yajia Huang verfasserin aut Mei Li verfasserin aut Lei Wang verfasserin aut Bing Li verfasserin aut Aiguo Xia verfasserin aut Ye Li verfasserin aut Shuai Yang verfasserin aut Fan Jin verfasserin aut In mLife Wiley, 2022 2(2023), 4, Seite 450-461 (DE-627)1804448230 (DE-600)3120417-X 2770100X nnns volume:2 year:2023 number:4 pages:450-461 https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/article/21f12515f24747119dd1548341552373 kostenfrei https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/toc/2770-100X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2023 4 450-461
allfields_unstemmed	10.1002/mlf2.12096 doi (DE-627)DOAJ097981982 (DE-599)DOAJ21f12515f24747119dd1548341552373 DE-627 ger DE-627 rakwb eng QR1-502 Rongrong Zhang verfasserin aut High‐throughput, microscopy‐based screening and quantification of genetic elements 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs. characterization methods high‐throughput robust genetic elements synthetic biology Microbiology Yajia Huang verfasserin aut Mei Li verfasserin aut Lei Wang verfasserin aut Bing Li verfasserin aut Aiguo Xia verfasserin aut Ye Li verfasserin aut Shuai Yang verfasserin aut Fan Jin verfasserin aut In mLife Wiley, 2022 2(2023), 4, Seite 450-461 (DE-627)1804448230 (DE-600)3120417-X 2770100X nnns volume:2 year:2023 number:4 pages:450-461 https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/article/21f12515f24747119dd1548341552373 kostenfrei https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/toc/2770-100X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2023 4 450-461
allfieldsGer	10.1002/mlf2.12096 doi (DE-627)DOAJ097981982 (DE-599)DOAJ21f12515f24747119dd1548341552373 DE-627 ger DE-627 rakwb eng QR1-502 Rongrong Zhang verfasserin aut High‐throughput, microscopy‐based screening and quantification of genetic elements 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs. characterization methods high‐throughput robust genetic elements synthetic biology Microbiology Yajia Huang verfasserin aut Mei Li verfasserin aut Lei Wang verfasserin aut Bing Li verfasserin aut Aiguo Xia verfasserin aut Ye Li verfasserin aut Shuai Yang verfasserin aut Fan Jin verfasserin aut In mLife Wiley, 2022 2(2023), 4, Seite 450-461 (DE-627)1804448230 (DE-600)3120417-X 2770100X nnns volume:2 year:2023 number:4 pages:450-461 https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/article/21f12515f24747119dd1548341552373 kostenfrei https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/toc/2770-100X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2023 4 450-461
allfieldsSound	10.1002/mlf2.12096 doi (DE-627)DOAJ097981982 (DE-599)DOAJ21f12515f24747119dd1548341552373 DE-627 ger DE-627 rakwb eng QR1-502 Rongrong Zhang verfasserin aut High‐throughput, microscopy‐based screening and quantification of genetic elements 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs. characterization methods high‐throughput robust genetic elements synthetic biology Microbiology Yajia Huang verfasserin aut Mei Li verfasserin aut Lei Wang verfasserin aut Bing Li verfasserin aut Aiguo Xia verfasserin aut Ye Li verfasserin aut Shuai Yang verfasserin aut Fan Jin verfasserin aut In mLife Wiley, 2022 2(2023), 4, Seite 450-461 (DE-627)1804448230 (DE-600)3120417-X 2770100X nnns volume:2 year:2023 number:4 pages:450-461 https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/article/21f12515f24747119dd1548341552373 kostenfrei https://doi.org/10.1002/mlf2.12096 kostenfrei https://doaj.org/toc/2770-100X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2023 4 450-461
language	English
source	In mLife 2(2023), 4, Seite 450-461 volume:2 year:2023 number:4 pages:450-461
sourceStr	In mLife 2(2023), 4, Seite 450-461 volume:2 year:2023 number:4 pages:450-461
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	characterization methods high‐throughput robust genetic elements synthetic biology Microbiology
isfreeaccess_bool	true
container_title	mLife
authorswithroles_txt_mv	Rongrong Zhang @@aut@@ Yajia Huang @@aut@@ Mei Li @@aut@@ Lei Wang @@aut@@ Bing Li @@aut@@ Aiguo Xia @@aut@@ Ye Li @@aut@@ Shuai Yang @@aut@@ Fan Jin @@aut@@
publishDateDaySort_date	2023-01-01T00:00:00Z
hierarchy_top_id	1804448230
id	DOAJ097981982
language_de	englisch
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callnumber-first	Q - Science
author	Rongrong Zhang
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topic_title	QR1-502 High‐throughput, microscopy‐based screening and quantification of genetic elements characterization methods high‐throughput robust genetic elements synthetic biology
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title	High‐throughput, microscopy‐based screening and quantification of genetic elements
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title_full	High‐throughput, microscopy‐based screening and quantification of genetic elements
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author_browse	Rongrong Zhang Yajia Huang Mei Li Lei Wang Bing Li Aiguo Xia Ye Li Shuai Yang Fan Jin
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title_sort	high‐throughput, microscopy‐based screening and quantification of genetic elements
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title_auth	High‐throughput, microscopy‐based screening and quantification of genetic elements
abstract	Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs.
abstractGer	Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs.
abstract_unstemmed	Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs.
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title_short	High‐throughput, microscopy‐based screening and quantification of genetic elements
url	https://doi.org/10.1002/mlf2.12096 https://doaj.org/article/21f12515f24747119dd1548341552373 https://doaj.org/toc/2770-100X
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High‐throughput, microscopy‐based screening and quantification of genetic elements

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