Spatiotemporal control of genome engineering in cone photoreceptors

Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the...
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Autor*in:	Wang, Nan-Kai [verfasserIn] Liu, Pei-Kang Kong, Yang Tseng, Yun-Ju Jenny, Laura A. Nolan, Nicholas D. Chen, Nelson Wang, Hung-Hsi Hsu, Chun Wei Huang, Wan-Chun Sparrow, Janet R. Lin, Chyuan-Sheng Tsang, Stephen H.

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2023

Schlagwörter:	Cone arrestin Arrestin 3 Cre-LoxP recombination CreERT2 2A peptide

Anmerkung:	© The Author(s) 2023. corrected publication 2023

Übergeordnetes Werk:	Enthalten in: Cell & bioscience - London : BioMed Central, 2011, 13(2023), 1 vom: 28. Juni
Übergeordnetes Werk:	volume:13 ; year:2023 ; number:1 ; day:28 ; month:06

Links:	Volltext

DOI / URN:	10.1186/s13578-023-01033-3

Katalog-ID:	SPR05208812X

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520			\|a Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models.
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allfields	10.1186/s13578-023-01033-3 doi (DE-627)SPR05208812X (SPR)s13578-023-01033-3-e DE-627 ger DE-627 rakwb eng Wang, Nan-Kai verfasserin aut Spatiotemporal control of genome engineering in cone photoreceptors 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s) 2023. corrected publication 2023 Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models. Cone arrestin (dpeaa)DE-He213 Arrestin 3 (dpeaa)DE-He213 Cre-LoxP recombination (dpeaa)DE-He213 CreERT2 (dpeaa)DE-He213 2A peptide (dpeaa)DE-He213 Liu, Pei-Kang aut Kong, Yang aut Tseng, Yun-Ju aut Jenny, Laura A. aut Nolan, Nicholas D. aut Chen, Nelson aut Wang, Hung-Hsi aut Hsu, Chun Wei aut Huang, Wan-Chun aut Sparrow, Janet R. aut Lin, Chyuan-Sheng aut Tsang, Stephen H. aut Enthalten in Cell & bioscience London : BioMed Central, 2011 13(2023), 1 vom: 28. Juni (DE-627)646079387 (DE-600)2593367-X 2045-3701 nnns volume:13 year:2023 number:1 day:28 month:06 https://dx.doi.org/10.1186/s13578-023-01033-3 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_11 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1 28 06
spelling	10.1186/s13578-023-01033-3 doi (DE-627)SPR05208812X (SPR)s13578-023-01033-3-e DE-627 ger DE-627 rakwb eng Wang, Nan-Kai verfasserin aut Spatiotemporal control of genome engineering in cone photoreceptors 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s) 2023. corrected publication 2023 Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models. Cone arrestin (dpeaa)DE-He213 Arrestin 3 (dpeaa)DE-He213 Cre-LoxP recombination (dpeaa)DE-He213 CreERT2 (dpeaa)DE-He213 2A peptide (dpeaa)DE-He213 Liu, Pei-Kang aut Kong, Yang aut Tseng, Yun-Ju aut Jenny, Laura A. aut Nolan, Nicholas D. aut Chen, Nelson aut Wang, Hung-Hsi aut Hsu, Chun Wei aut Huang, Wan-Chun aut Sparrow, Janet R. aut Lin, Chyuan-Sheng aut Tsang, Stephen H. aut Enthalten in Cell & bioscience London : BioMed Central, 2011 13(2023), 1 vom: 28. Juni (DE-627)646079387 (DE-600)2593367-X 2045-3701 nnns volume:13 year:2023 number:1 day:28 month:06 https://dx.doi.org/10.1186/s13578-023-01033-3 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_11 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1 28 06
allfields_unstemmed	10.1186/s13578-023-01033-3 doi (DE-627)SPR05208812X (SPR)s13578-023-01033-3-e DE-627 ger DE-627 rakwb eng Wang, Nan-Kai verfasserin aut Spatiotemporal control of genome engineering in cone photoreceptors 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s) 2023. corrected publication 2023 Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models. Cone arrestin (dpeaa)DE-He213 Arrestin 3 (dpeaa)DE-He213 Cre-LoxP recombination (dpeaa)DE-He213 CreERT2 (dpeaa)DE-He213 2A peptide (dpeaa)DE-He213 Liu, Pei-Kang aut Kong, Yang aut Tseng, Yun-Ju aut Jenny, Laura A. aut Nolan, Nicholas D. aut Chen, Nelson aut Wang, Hung-Hsi aut Hsu, Chun Wei aut Huang, Wan-Chun aut Sparrow, Janet R. aut Lin, Chyuan-Sheng aut Tsang, Stephen H. aut Enthalten in Cell & bioscience London : BioMed Central, 2011 13(2023), 1 vom: 28. Juni (DE-627)646079387 (DE-600)2593367-X 2045-3701 nnns volume:13 year:2023 number:1 day:28 month:06 https://dx.doi.org/10.1186/s13578-023-01033-3 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_11 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1 28 06
allfieldsGer	10.1186/s13578-023-01033-3 doi (DE-627)SPR05208812X (SPR)s13578-023-01033-3-e DE-627 ger DE-627 rakwb eng Wang, Nan-Kai verfasserin aut Spatiotemporal control of genome engineering in cone photoreceptors 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s) 2023. corrected publication 2023 Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models. Cone arrestin (dpeaa)DE-He213 Arrestin 3 (dpeaa)DE-He213 Cre-LoxP recombination (dpeaa)DE-He213 CreERT2 (dpeaa)DE-He213 2A peptide (dpeaa)DE-He213 Liu, Pei-Kang aut Kong, Yang aut Tseng, Yun-Ju aut Jenny, Laura A. aut Nolan, Nicholas D. aut Chen, Nelson aut Wang, Hung-Hsi aut Hsu, Chun Wei aut Huang, Wan-Chun aut Sparrow, Janet R. aut Lin, Chyuan-Sheng aut Tsang, Stephen H. aut Enthalten in Cell & bioscience London : BioMed Central, 2011 13(2023), 1 vom: 28. Juni (DE-627)646079387 (DE-600)2593367-X 2045-3701 nnns volume:13 year:2023 number:1 day:28 month:06 https://dx.doi.org/10.1186/s13578-023-01033-3 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_11 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1 28 06
allfieldsSound	10.1186/s13578-023-01033-3 doi (DE-627)SPR05208812X (SPR)s13578-023-01033-3-e DE-627 ger DE-627 rakwb eng Wang, Nan-Kai verfasserin aut Spatiotemporal control of genome engineering in cone photoreceptors 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s) 2023. corrected publication 2023 Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models. Cone arrestin (dpeaa)DE-He213 Arrestin 3 (dpeaa)DE-He213 Cre-LoxP recombination (dpeaa)DE-He213 CreERT2 (dpeaa)DE-He213 2A peptide (dpeaa)DE-He213 Liu, Pei-Kang aut Kong, Yang aut Tseng, Yun-Ju aut Jenny, Laura A. aut Nolan, Nicholas D. aut Chen, Nelson aut Wang, Hung-Hsi aut Hsu, Chun Wei aut Huang, Wan-Chun aut Sparrow, Janet R. aut Lin, Chyuan-Sheng aut Tsang, Stephen H. aut Enthalten in Cell & bioscience London : BioMed Central, 2011 13(2023), 1 vom: 28. Juni (DE-627)646079387 (DE-600)2593367-X 2045-3701 nnns volume:13 year:2023 number:1 day:28 month:06 https://dx.doi.org/10.1186/s13578-023-01033-3 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_11 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1 28 06
language	English
source	Enthalten in Cell & bioscience 13(2023), 1 vom: 28. Juni volume:13 year:2023 number:1 day:28 month:06
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topic_title	Spatiotemporal control of genome engineering in cone photoreceptors Cone arrestin (dpeaa)DE-He213 Arrestin 3 (dpeaa)DE-He213 Cre-LoxP recombination (dpeaa)DE-He213 CreERT2 (dpeaa)DE-He213 2A peptide (dpeaa)DE-He213
topic	misc Cone arrestin misc Arrestin 3 misc Cre-LoxP recombination misc CreERT2 misc 2A peptide
topic_unstemmed	misc Cone arrestin misc Arrestin 3 misc Cre-LoxP recombination misc CreERT2 misc 2A peptide
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title_sort	spatiotemporal control of genome engineering in cone photoreceptors
title_auth	Spatiotemporal control of genome engineering in cone photoreceptors
abstract	Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models. © The Author(s) 2023. corrected publication 2023
abstractGer	Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models. © The Author(s) 2023. corrected publication 2023
abstract_unstemmed	Background Cones are essential for color recognition, high resolution, and central vision; therefore cone death causes blindness. Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. Moreover, the Cre activity can be induced by delivering tamoxifen intragastrically as early as PD2, which will be useful for studying retinal development or in rapid degenerative mouse models. © The Author(s) 2023. corrected publication 2023
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title_short	Spatiotemporal control of genome engineering in cone photoreceptors
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Understanding the pathophysiology of each cell type in the retina is key to developing therapies for retinal diseases. However, studying the biology of cone cells in the rod-dominant mammalian retina is particularly challenging. In this study, we used a bacterial artificial chromosome (BAC) recombineering method to knock in the “$ CreER^{T2} $” sequence into the Gnat2 and Arr3 genes, respectively and generated three novel inducible $ CreER^{T2} $ mice with different cone cell specificities. Results These models (Gnat2CreERT2, Arr3T2ACreERT2, and Arr3P2ACreERT2) express temporally controllable Cre recombinase that achieves conditional alleles in cone photoreceptors. Cre-LoxP recombination can be induced as early as postnatal day (PD) two upon tamoxifen injection at varying efficiencies, ranging from 10 to 15% in Gnat2CreERT2, 40% in Arr3T2ACreERT2, and 100% in Arr3P2ACreERT2. Notably, knocking in the P2A-CreERT2 cassette does not affect cone cell morphology and functionality. Most cone-phototransduction enzymes, including Opsins, CNGA3, etc. are not altered except for a reduction in the Arr3 transcript. Conclusions The Arr3P2ACreERT2 mouse, an inducible cone-specific Cre driver, is a valuable line in studying cone cell biology, function, as well as its relationship with rod and other retinal cells. 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Spatiotemporal control of genome engineering in cone photoreceptors

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