DNA Repair Pathway Choices in CRISPR-Cas9-Mediated Genome Editing

Many clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based genome editing technologies take advantage of Cas nucleases to induce DNA double-strand breaks (DSBs) at desired locations within a genome. Further processing of the DSBs by the cellular...

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Bibliographic Details
Published inTrends in genetics Vol. 37; no. 7; pp. 639 - 656
Main Authors Xue, Chaoyou, Greene, Eric C.
Format Journal Article
LanguageEnglish
Published England Elsevier Ltd 01.07.2021
Subjects
CRISPR-Cas Systems - genetics
DNA Breaks, Double-Stranded
DNA End-Joining Repair - genetics
DNA Repair - genetics
Gene Editing - trends
Genome, Human - genetics
Homologous Recombination - genetics
Humans
Mutagenesis, Insertional - genetics
Signal Transduction - genetics
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Abstract Many clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based genome editing technologies take advantage of Cas nucleases to induce DNA double-strand breaks (DSBs) at desired locations within a genome. Further processing of the DSBs by the cellular DSB repair machinery is then necessary to introduce desired mutations, sequence insertions, or gene deletions. Thus, the accuracy and efficiency of genome editing are influenced by the cellular DSB repair pathways. DSBs are themselves highly genotoxic lesions and as such cells have evolved multiple mechanisms for their repair. These repair pathways include homologous recombination (HR), classical nonhomologous end joining (cNHEJ), microhomology-mediated end joining (MMEJ) and single-strand annealing (SSA). In this review, we briefly highlight CRISPR-Cas9 and then describe the mechanisms of DSB repair. Finally, we summarize recent findings of factors that can influence the choice of DNA repair pathway in response to Cas9-induced DSBs. Clustered regularly interspaced short palindromic repeats (CRISPR)- CRISPR-associated protein 9 (Cas9)-mediated genome editing offers a powerful approach as a potential therapy for monogenic human genetic diseases.Precise template-free base deletions can be achieved through microhomology-mediated end joining (MMEJ) repair and depend on local target site sequence.The DNA repair pathway choice in CRISPR-Cas9 induced-double-strand breaks (DSBs) is regulated by several key factors including the cell cycle, target site sequence and chromatin structure, and the identity of the donor DNA template.Homology-directed repair (HDR)-related DNA repair pathways in response to CRISPR-Cas9 induced-DSBs in mammalian cells are complicated and relatively inefficient. Different DNA repair pathways might be used to repair each end at a DSB resulting in the potential for asymmetric repair.
AbstractList Many clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based genome editing technologies take advantage of Cas nucleases to induce DNA double-strand breaks (DSBs) at desired locations within a genome. Further processing of the DSBs by the cellular DSB repair machinery is then necessary to introduce desired mutations, sequence insertions, or gene deletions. Thus, the accuracy and efficiency of genome editing are influenced by the cellular DSB repair pathways. DSBs are themselves highly genotoxic lesions and as such cells have evolved multiple mechanisms for their repair. These repair pathways include homologous recombination (HR), classical nonhomologous end joining (cNHEJ), microhomology-mediated end joining (MMEJ) and single-strand annealing (SSA). In this review, we briefly highlight CRISPR-Cas9 and then describe the mechanisms of DSB repair. Finally, we summarize recent findings of factors that can influence the choice of DNA repair pathway in response to Cas9-induced DSBs.
Many clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based genome editing technologies take advantage of Cas nucleases to induce DNA double-strand breaks (DSBs) at desired locations within a genome. Further processing of the DSBs by the cellular DSB repair machinery is then necessary to introduce desired mutations, sequence insertions, or gene deletions. Thus, the accuracy and efficiency of genome editing are influenced by the cellular DSB repair pathways. DSBs are themselves highly genotoxic lesions and as such cells have evolved multiple mechanisms for their repair. These repair pathways include homologous recombination (HR), classical nonhomologous end joining (cNHEJ), microhomology-mediated end joining (MMEJ) and single-strand annealing (SSA). In this review, we briefly highlight CRISPR-Cas9 and then describe the mechanisms of DSB repair. Finally, we summarize recent findings of factors that can influence the choice of DNA repair pathway in response to Cas9-induced DSBs. Clustered regularly interspaced short palindromic repeats (CRISPR)- CRISPR-associated protein 9 (Cas9)-mediated genome editing offers a powerful approach as a potential therapy for monogenic human genetic diseases.Precise template-free base deletions can be achieved through microhomology-mediated end joining (MMEJ) repair and depend on local target site sequence.The DNA repair pathway choice in CRISPR-Cas9 induced-double-strand breaks (DSBs) is regulated by several key factors including the cell cycle, target site sequence and chromatin structure, and the identity of the donor DNA template.Homology-directed repair (HDR)-related DNA repair pathways in response to CRISPR-Cas9 induced-DSBs in mammalian cells are complicated and relatively inefficient. Different DNA repair pathways might be used to repair each end at a DSB resulting in the potential for asymmetric repair.
Many CRISPR-Cas-based genome editing technologies take advantage of Cas nucleases to induce DNA double-strand breaks (DSBs) at desired locations within a genome. Further processing of the DSBs by the cellular DSB repair machinery is then necessary to introduce desired mutations, sequence insertions, or gene deletions. Thus, the accuracy and efficiency of genome editing are influenced by the cellular DSB repair pathways. DSBs are themselves highly genotoxic lesions and as such cells have evolved multiple mechanisms for their repair. These repair pathways include homologous recombination (HR), classical non-homologous end joining (cNHEJ), microhomology-mediated end joining (MMEJ) and single-strand annealing (SSA). In this review, we briefly highlight the CRISPR-Cas9 system and we then describe the mechanisms of DSB repair. Finally, we summarize recent findings of factors that can influence the choice of DNA repair pathway response to Cas9-induced DSBs.
Many clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based genome editing technologies take advantage of Cas nucleases to induce DNA double-strand breaks (DSBs) at desired locations within a genome. Further processing of the DSBs by the cellular DSB repair machinery is then necessary to introduce desired mutations, sequence insertions, or gene deletions. Thus, the accuracy and efficiency of genome editing are influenced by the cellular DSB repair pathways. DSBs are themselves highly genotoxic lesions and as such cells have evolved multiple mechanisms for their repair. These repair pathways include homologous recombination (HR), classical nonhomologous end joining (cNHEJ), microhomology-mediated end joining (MMEJ) and single-strand annealing (SSA). In this review, we briefly highlight CRISPR-Cas9 and then describe the mechanisms of DSB repair. Finally, we summarize recent findings of factors that can influence the choice of DNA repair pathway in response to Cas9-induced DSBs.Many clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based genome editing technologies take advantage of Cas nucleases to induce DNA double-strand breaks (DSBs) at desired locations within a genome. Further processing of the DSBs by the cellular DSB repair machinery is then necessary to introduce desired mutations, sequence insertions, or gene deletions. Thus, the accuracy and efficiency of genome editing are influenced by the cellular DSB repair pathways. DSBs are themselves highly genotoxic lesions and as such cells have evolved multiple mechanisms for their repair. These repair pathways include homologous recombination (HR), classical nonhomologous end joining (cNHEJ), microhomology-mediated end joining (MMEJ) and single-strand annealing (SSA). In this review, we briefly highlight CRISPR-Cas9 and then describe the mechanisms of DSB repair. Finally, we summarize recent findings of factors that can influence the choice of DNA repair pathway in response to Cas9-induced DSBs.
Author Xue, Chaoyou
Greene, Eric C.
AuthorAffiliation 1 Department of Biochemistry & Molecular Biophysics, Columbia University, New York, NY, 10032
AuthorAffiliation_xml – name: 1 Department of Biochemistry & Molecular Biophysics, Columbia University, New York, NY, 10032
Author_xml – sequence: 1
  givenname: Chaoyou
  surname: Xue
  fullname: Xue, Chaoyou
  organization: Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA
– sequence: 2
  givenname: Eric C.
  surname: Greene
  fullname: Greene, Eric C.
  email: ecg2108@cumc.columbia.edu
  organization: Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA
BackLink https://www.ncbi.nlm.nih.gov/pubmed/33896583$$D View this record in MEDLINE/PubMed
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Present Address: Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
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PublicationTitle Trends in genetics
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Feng (bb0535) 2011; 108
Zetsche (bb0270) 2015; 163
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Jiang (bb0600) 2013; 4
Maruyama (bb0350) 2015; 33
Hu (bb0355) 2018; 8
Yang (bb0155) 2016; 34
Ramakrishna (bb0160) 2014; 24
Clouaire, Legube (bb0870) 2019; 35
Ishino (bb0945) 1987; 169
van Overbeek (10.1016/j.tig.2021.02.008_bb0495) 2016; 63
Lee (10.1016/j.tig.2021.02.008_bb0125) 2018; 9
Wang (10.1016/j.tig.2021.02.008_bb0195) 2016; 113
Lamarche (10.1016/j.tig.2021.02.008_bb0580) 2010; 584
Hu (10.1016/j.tig.2021.02.008_bb0355) 2018; 8
Crickard (10.1016/j.tig.2021.02.008_bb0775) 2020; 181
Chen (10.1016/j.tig.2021.02.008_bb0070) 2014; 289
Eid (10.1016/j.tig.2021.02.008_bb0635) 2010; 11
Sartori (10.1016/j.tig.2021.02.008_bb0415) 2007; 450
Shao (10.1016/j.tig.2021.02.008_bb0905) 2014; 9
Brouns (10.1016/j.tig.2021.02.008_bb0965) 2008; 321
Daley (10.1016/j.tig.2021.02.008_bb0655) 2014; 42
Chang (10.1016/j.tig.2021.02.008_bb0050) 2017; 18
Silva (10.1016/j.tig.2021.02.008_bb0205) 2011; 11
Mari (10.1016/j.tig.2021.02.008_bb0315) 2006; 103
Ronato (10.1016/j.tig.2021.02.008_bb0630) 2020; 45
Senis (10.1016/j.tig.2021.02.008_bb0140) 2014; 9
Murugan (10.1016/j.tig.2021.02.008_bb0285) 2020; 295
Schumann (10.1016/j.tig.2021.02.008_bb0210) 2015; 112
Fernandes-Alnemri (10.1016/j.tig.2021.02.008_bb0235) 2009; 458
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Chen (10.1016/j.tig.2021.02.008_bb0085) 2017; 550
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Deltcheva (10.1016/j.tig.2021.02.008_bb0990) 2011; 471
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Isaac (10.1016/j.tig.2021.02.008_bb0860) 2016; 5
Lou (10.1016/j.tig.2021.02.008_bb0685) 2003; 421
Kleinstiver (10.1016/j.tig.2021.02.008_bb0120) 2016; 529
Yang (10.1016/j.tig.2021.02.008_bb0755) 2002; 297
Yun (10.1016/j.tig.2021.02.008_bb0615) 2009; 459
Jinek (10.1016/j.tig.2021.02.008_bb0025) 2013; 2
Hendel (10.1016/j.tig.2021.02.008_bb0165) 2015; 33
Gutschner (10.1016/j.tig.2021.02.008_bb0845) 2016; 14
Kuscu (10.1016/j.tig.2021.02.008_bb0110) 2014; 32
Nick McElhinny (10.1016/j.tig.2021.02.008_bb0305) 2000; 20
Caron (10.1016/j.tig.2021.02.008_bb0460) 2019; 10
Lu (10.1016/j.tig.2021.02.008_bb0765) 2018; 115
Chen (10.1016/j.tig.2021.02.008_bb0710) 2008; 283
Wang (10.1016/j.tig.2021.02.008_bb0455) 2006; 34
Fu (10.1016/j.tig.2021.02.008_bb0105) 2013; 31
Ishino (10.1016/j.tig.2021.02.008_bb0945) 1987; 169
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Tarsounas (10.1016/j.tig.2021.02.008_bb0695) 2020; 21
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Audebert (10.1016/j.tig.2021.02.008_bb0450) 2004; 279
Chen (10.1016/j.tig.2021.02.008_bb0725) 2018; 2
Anzalone (10.1016/j.tig.2021.02.008_bb0265) 2020; 38
Yoshimi (10.1016/j.tig.2021.02.008_bb0900) 2016; 7
Kim (10.1016/j.tig.2021.02.008_bb0280) 2016; 34
Wright (10.1016/j.tig.2021.02.008_bb0995) 2016; 164
Aylon (10.1016/j.tig.2021.02.008_bb0830) 2004; 23
Myler (10.1016/j.tig.2021.02.008_bb0425) 2017; 67
Daley (10.1016/j.tig.2021.02.008_bb0660) 2017; 21
Prakash (10.1016/j.tig.2021.02.008_bb0735) 2020
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Deveryshetty (10.1016/j.tig.2021.02.008_bb0800) 2019; 8
Wang (10.1016/j.tig.2021.02.008_bb0345) 2020; 79
Chu (10.1016/j.tig.2021.02.008_bb0360) 2015; 33
Xia (10.1016/j.tig.2021.02.008_bb0745) 2006; 22
Liang (10.1016/j.tig.2021.02.008_bb0390) 2008; 36
Allen (10.1016/j.tig.2021.02.008_bb0490) 2018
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Bhargava (10.1016/j.tig.2021.02.008_bb0060) 2016; 32
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He (10.1016/j.tig.2021.02.008_bb0620) 2018; 563
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Shen (10.1016/j.tig.2021.02.008_bb0480) 2018; 563
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Sturzenegger (10.1016/j.tig.2021.02.008_bb0505) 2014; 289
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Shrivastav (10.1016/j.tig.2021.02.008_bb0560) 2008; 18
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Haince (10.1016/j.tig.2021.02.008_bb0590) 2008; 283
Liang (10.1016/j.tig.2021.02.008_bb0785) 2016; 15
Doudna (10.1016/j.tig.2021.02.008_bb0005) 2014; 346
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Miller (10.1016/j.tig.2021.02.008_bb0015) 2011; 29
Pattanayak (10.1016/j.tig.2021.02.008_bb0100) 2013; 31
Srivastava (10.1016/j.tig.2021.02.008_bb0435) 2012; 151
Meek (10.1016/j.tig.2021.02.008_bb0300) 2008; 99
Vakulskas (10.1016/j.tig.2021.02.008_bb0130) 2018; 24
Seol (10.1016/j.tig.2021.02.008_bb0380) 2018; 809
Bunting (10.1016/j.tig.2021.02.008_bb0700) 2010; 141
Sung (10.1016/j.tig.2021.02.008_bb0530) 1997; 272
Jinek (10.1016/j.tig.2021.02.008_bb0020) 2012; 337
Grimme (10.1016/j.tig.2021.02.008_bb0520) 2010; 38
Cong (10.1016/j.tig.2021.02.008_bb0030) 2013; 339
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Dray (10.1016/j.tig.2021.02.008_bb0790) 2010; 17
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Lino (10.1016/j.tig.2021.02.008_bb0245) 2018; 25
Zhou (10.1016/j.tig.2021.02.008_bb0650) 2015; 4
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Chen (10.1016/j.tig.2021.02.008_bb0555) 2013; 50
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Yang (10.1016/j.tig.2021.02.008_bb0155) 2016; 34
Chu (10.1016/j.tig.2021.02.008_bb0770) 2015; 6
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Yin (10.1016/j.tig.2021.02.008_bb0135) 2017; 16
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Snippet Many clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based genome editing technologies take advantage of...
Many CRISPR-Cas-based genome editing technologies take advantage of Cas nucleases to induce DNA double-strand breaks (DSBs) at desired locations within a...
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SubjectTerms CRISPR-Cas Systems - genetics
DNA Breaks, Double-Stranded
DNA End-Joining Repair - genetics
DNA Repair - genetics
Gene Editing - trends
Genome, Human - genetics
Homologous Recombination - genetics
Humans
Mutagenesis, Insertional - genetics
Signal Transduction - genetics
Title DNA Repair Pathway Choices in CRISPR-Cas9-Mediated Genome Editing
URI https://dx.doi.org/10.1016/j.tig.2021.02.008
https://www.ncbi.nlm.nih.gov/pubmed/33896583
https://www.proquest.com/docview/2518735683
https://pubmed.ncbi.nlm.nih.gov/PMC8187289
Volume 37
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