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5′-Modifications improve potency and efficacy of DNA donors for precision genome editing

Nuclease-directed genome editing is a powerful tool for investigating physiology and has great promise as a therapeutic approach to correct mutations that cause disease. In its most precise form, genome editing can use cellular homology-directed repair (HDR) pathways to insert information from an ex...

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Autores principales: Ghanta, Krishna S, Chen, Zexiang, Mir, Aamir, Dokshin, Gregoriy A, Krishnamurthy, Pranathi M, Yoon, Yeonsoo, Gallant, Judith, Xu, Ping, Zhang, Xiao-Ou, Ozturk, Ahmet Rasit, Shin, Masahiro, Idrizi, Feston, Liu, Pengpeng, Gneid, Hassan, Edraki, Alireza, Lawson, Nathan D, Rivera-Pérez, Jaime A, Sontheimer, Erik J, Watts, Jonathan K, Mello, Craig C
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8568340/
https://www.ncbi.nlm.nih.gov/pubmed/34665130
http://dx.doi.org/10.7554/eLife.72216
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author Ghanta, Krishna S
Chen, Zexiang
Mir, Aamir
Dokshin, Gregoriy A
Krishnamurthy, Pranathi M
Yoon, Yeonsoo
Gallant, Judith
Xu, Ping
Zhang, Xiao-Ou
Ozturk, Ahmet Rasit
Shin, Masahiro
Idrizi, Feston
Liu, Pengpeng
Gneid, Hassan
Edraki, Alireza
Lawson, Nathan D
Rivera-Pérez, Jaime A
Sontheimer, Erik J
Watts, Jonathan K
Mello, Craig C
author_facet Ghanta, Krishna S
Chen, Zexiang
Mir, Aamir
Dokshin, Gregoriy A
Krishnamurthy, Pranathi M
Yoon, Yeonsoo
Gallant, Judith
Xu, Ping
Zhang, Xiao-Ou
Ozturk, Ahmet Rasit
Shin, Masahiro
Idrizi, Feston
Liu, Pengpeng
Gneid, Hassan
Edraki, Alireza
Lawson, Nathan D
Rivera-Pérez, Jaime A
Sontheimer, Erik J
Watts, Jonathan K
Mello, Craig C
author_sort Ghanta, Krishna S
collection PubMed
description Nuclease-directed genome editing is a powerful tool for investigating physiology and has great promise as a therapeutic approach to correct mutations that cause disease. In its most precise form, genome editing can use cellular homology-directed repair (HDR) pathways to insert information from an exogenously supplied DNA-repair template (donor) directly into a targeted genomic location. Unfortunately, particularly for long insertions, toxicity and delivery considerations associated with repair template DNA can limit HDR efficacy. Here, we explore chemical modifications to both double-stranded and single-stranded DNA-repair templates. We describe 5′-terminal modifications, including in its simplest form the incorporation of triethylene glycol (TEG) moieties, that consistently increase the frequency of precision editing in the germlines of three animal models (Caenorhabditis elegans, zebrafish, mice) and in cultured human cells.
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spelling pubmed-85683402021-11-08 5′-Modifications improve potency and efficacy of DNA donors for precision genome editing Ghanta, Krishna S Chen, Zexiang Mir, Aamir Dokshin, Gregoriy A Krishnamurthy, Pranathi M Yoon, Yeonsoo Gallant, Judith Xu, Ping Zhang, Xiao-Ou Ozturk, Ahmet Rasit Shin, Masahiro Idrizi, Feston Liu, Pengpeng Gneid, Hassan Edraki, Alireza Lawson, Nathan D Rivera-Pérez, Jaime A Sontheimer, Erik J Watts, Jonathan K Mello, Craig C eLife Biochemistry and Chemical Biology Nuclease-directed genome editing is a powerful tool for investigating physiology and has great promise as a therapeutic approach to correct mutations that cause disease. In its most precise form, genome editing can use cellular homology-directed repair (HDR) pathways to insert information from an exogenously supplied DNA-repair template (donor) directly into a targeted genomic location. Unfortunately, particularly for long insertions, toxicity and delivery considerations associated with repair template DNA can limit HDR efficacy. Here, we explore chemical modifications to both double-stranded and single-stranded DNA-repair templates. We describe 5′-terminal modifications, including in its simplest form the incorporation of triethylene glycol (TEG) moieties, that consistently increase the frequency of precision editing in the germlines of three animal models (Caenorhabditis elegans, zebrafish, mice) and in cultured human cells. eLife Sciences Publications, Ltd 2021-10-19 /pmc/articles/PMC8568340/ /pubmed/34665130 http://dx.doi.org/10.7554/eLife.72216 Text en © 2021, Ghanta et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biochemistry and Chemical Biology
Ghanta, Krishna S
Chen, Zexiang
Mir, Aamir
Dokshin, Gregoriy A
Krishnamurthy, Pranathi M
Yoon, Yeonsoo
Gallant, Judith
Xu, Ping
Zhang, Xiao-Ou
Ozturk, Ahmet Rasit
Shin, Masahiro
Idrizi, Feston
Liu, Pengpeng
Gneid, Hassan
Edraki, Alireza
Lawson, Nathan D
Rivera-Pérez, Jaime A
Sontheimer, Erik J
Watts, Jonathan K
Mello, Craig C
5′-Modifications improve potency and efficacy of DNA donors for precision genome editing
title 5′-Modifications improve potency and efficacy of DNA donors for precision genome editing
title_full 5′-Modifications improve potency and efficacy of DNA donors for precision genome editing
title_fullStr 5′-Modifications improve potency and efficacy of DNA donors for precision genome editing
title_full_unstemmed 5′-Modifications improve potency and efficacy of DNA donors for precision genome editing
title_short 5′-Modifications improve potency and efficacy of DNA donors for precision genome editing
title_sort 5′-modifications improve potency and efficacy of dna donors for precision genome editing
topic Biochemistry and Chemical Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8568340/
https://www.ncbi.nlm.nih.gov/pubmed/34665130
http://dx.doi.org/10.7554/eLife.72216
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