Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts
The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals wi...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Li, Guoling [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2018transfer abstract |
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| Schlagwörter: |
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| Umfang: |
7 |
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| Übergeordnetes Werk: |
Enthalten in: Urological Diseases of the Byzantine Emperors (330-1453) - 2011, Amsterdam |
|---|---|
| Übergeordnetes Werk: |
volume:99 ; year:2018 ; pages:154-160 ; extent:7 |
| Links: |
|---|
| DOI / URN: |
10.1016/j.biocel.2018.04.011 |
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ELV043063101 |
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| 245 | 1 | 0 | |a Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts |
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| 520 | |a The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. | ||
| 520 | |a The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. | ||
| 650 | 7 | |a Ku70/80 |2 Elsevier | |
| 650 | 7 | |a RNA interference |2 Elsevier | |
| 650 | 7 | |a Genome editing |2 Elsevier | |
| 650 | 7 | |a Pig fetal fibroblasts |2 Elsevier | |
| 650 | 7 | |a Homology-directed repair |2 Elsevier | |
| 700 | 1 | |a Liu, Dewu |4 oth | |
| 700 | 1 | |a Zhang, Xianwei |4 oth | |
| 700 | 1 | |a Quan, Rong |4 oth | |
| 700 | 1 | |a Zhong, Cuili |4 oth | |
| 700 | 1 | |a Mo, Jianxin |4 oth | |
| 700 | 1 | |a Huang, Yaoqiang |4 oth | |
| 700 | 1 | |a Wang, Haoqiang |4 oth | |
| 700 | 1 | |a Ruan, Xiaofang |4 oth | |
| 700 | 1 | |a Xu, Zheng |4 oth | |
| 700 | 1 | |a Zheng, Enqin |4 oth | |
| 700 | 1 | |a Gu, Ting |4 oth | |
| 700 | 1 | |a Hong, Linjun |4 oth | |
| 700 | 1 | |a Li, Zicong |4 oth | |
| 700 | 1 | |a Wu, Zhenfang |4 oth | |
| 700 | 1 | |a Yang, Huaqiang |4 oth | |
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10.1016/j.biocel.2018.04.011 doi GBV00000000000619.pica (DE-627)ELV043063101 (ELSEVIER)S1357-2725(18)30086-4 DE-627 ger DE-627 rakwb eng 610 VZ 610 VZ 44.85 bkl Li, Guoling verfasserin aut Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts 2018transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. Ku70/80 Elsevier RNA interference Elsevier Genome editing Elsevier Pig fetal fibroblasts Elsevier Homology-directed repair Elsevier Liu, Dewu oth Zhang, Xianwei oth Quan, Rong oth Zhong, Cuili oth Mo, Jianxin oth Huang, Yaoqiang oth Wang, Haoqiang oth Ruan, Xiaofang oth Xu, Zheng oth Zheng, Enqin oth Gu, Ting oth Hong, Linjun oth Li, Zicong oth Wu, Zhenfang oth Yang, Huaqiang oth Enthalten in Elsevier Urological Diseases of the Byzantine Emperors (330-1453) 2011 Amsterdam (DE-627)ELV010616845 volume:99 year:2018 pages:154-160 extent:7 https://doi.org/10.1016/j.biocel.2018.04.011 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA 44.85 Kardiologie Angiologie VZ AR 99 2018 154-160 7 |
| spelling |
10.1016/j.biocel.2018.04.011 doi GBV00000000000619.pica (DE-627)ELV043063101 (ELSEVIER)S1357-2725(18)30086-4 DE-627 ger DE-627 rakwb eng 610 VZ 610 VZ 44.85 bkl Li, Guoling verfasserin aut Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts 2018transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. Ku70/80 Elsevier RNA interference Elsevier Genome editing Elsevier Pig fetal fibroblasts Elsevier Homology-directed repair Elsevier Liu, Dewu oth Zhang, Xianwei oth Quan, Rong oth Zhong, Cuili oth Mo, Jianxin oth Huang, Yaoqiang oth Wang, Haoqiang oth Ruan, Xiaofang oth Xu, Zheng oth Zheng, Enqin oth Gu, Ting oth Hong, Linjun oth Li, Zicong oth Wu, Zhenfang oth Yang, Huaqiang oth Enthalten in Elsevier Urological Diseases of the Byzantine Emperors (330-1453) 2011 Amsterdam (DE-627)ELV010616845 volume:99 year:2018 pages:154-160 extent:7 https://doi.org/10.1016/j.biocel.2018.04.011 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA 44.85 Kardiologie Angiologie VZ AR 99 2018 154-160 7 |
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10.1016/j.biocel.2018.04.011 doi GBV00000000000619.pica (DE-627)ELV043063101 (ELSEVIER)S1357-2725(18)30086-4 DE-627 ger DE-627 rakwb eng 610 VZ 610 VZ 44.85 bkl Li, Guoling verfasserin aut Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts 2018transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. Ku70/80 Elsevier RNA interference Elsevier Genome editing Elsevier Pig fetal fibroblasts Elsevier Homology-directed repair Elsevier Liu, Dewu oth Zhang, Xianwei oth Quan, Rong oth Zhong, Cuili oth Mo, Jianxin oth Huang, Yaoqiang oth Wang, Haoqiang oth Ruan, Xiaofang oth Xu, Zheng oth Zheng, Enqin oth Gu, Ting oth Hong, Linjun oth Li, Zicong oth Wu, Zhenfang oth Yang, Huaqiang oth Enthalten in Elsevier Urological Diseases of the Byzantine Emperors (330-1453) 2011 Amsterdam (DE-627)ELV010616845 volume:99 year:2018 pages:154-160 extent:7 https://doi.org/10.1016/j.biocel.2018.04.011 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA 44.85 Kardiologie Angiologie VZ AR 99 2018 154-160 7 |
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10.1016/j.biocel.2018.04.011 doi GBV00000000000619.pica (DE-627)ELV043063101 (ELSEVIER)S1357-2725(18)30086-4 DE-627 ger DE-627 rakwb eng 610 VZ 610 VZ 44.85 bkl Li, Guoling verfasserin aut Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts 2018transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. Ku70/80 Elsevier RNA interference Elsevier Genome editing Elsevier Pig fetal fibroblasts Elsevier Homology-directed repair Elsevier Liu, Dewu oth Zhang, Xianwei oth Quan, Rong oth Zhong, Cuili oth Mo, Jianxin oth Huang, Yaoqiang oth Wang, Haoqiang oth Ruan, Xiaofang oth Xu, Zheng oth Zheng, Enqin oth Gu, Ting oth Hong, Linjun oth Li, Zicong oth Wu, Zhenfang oth Yang, Huaqiang oth Enthalten in Elsevier Urological Diseases of the Byzantine Emperors (330-1453) 2011 Amsterdam (DE-627)ELV010616845 volume:99 year:2018 pages:154-160 extent:7 https://doi.org/10.1016/j.biocel.2018.04.011 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA 44.85 Kardiologie Angiologie VZ AR 99 2018 154-160 7 |
| allfieldsSound |
10.1016/j.biocel.2018.04.011 doi GBV00000000000619.pica (DE-627)ELV043063101 (ELSEVIER)S1357-2725(18)30086-4 DE-627 ger DE-627 rakwb eng 610 VZ 610 VZ 44.85 bkl Li, Guoling verfasserin aut Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts 2018transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. Ku70/80 Elsevier RNA interference Elsevier Genome editing Elsevier Pig fetal fibroblasts Elsevier Homology-directed repair Elsevier Liu, Dewu oth Zhang, Xianwei oth Quan, Rong oth Zhong, Cuili oth Mo, Jianxin oth Huang, Yaoqiang oth Wang, Haoqiang oth Ruan, Xiaofang oth Xu, Zheng oth Zheng, Enqin oth Gu, Ting oth Hong, Linjun oth Li, Zicong oth Wu, Zhenfang oth Yang, Huaqiang oth Enthalten in Elsevier Urological Diseases of the Byzantine Emperors (330-1453) 2011 Amsterdam (DE-627)ELV010616845 volume:99 year:2018 pages:154-160 extent:7 https://doi.org/10.1016/j.biocel.2018.04.011 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA 44.85 Kardiologie Angiologie VZ AR 99 2018 154-160 7 |
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suppressing ku70/ku80 expression elevates homology-directed repair efficiency in primary fibroblasts |
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Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts |
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The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. |
| abstractGer |
The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. |
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The main DNA repair pathways, nonhomologous end joining (NHEJ) and homology-directed repair (HDR), are complementary to each other; hence, interruptions of the NHEJ pathway can favor HDR. Improving HDR efficiency in animal primary fibroblasts can facilitate the generation of gene knock-in animals with agricultural and biomedical values by somatic cell nuclear transfer. In this study, we used siRNA to suppress the expression of Ku70 and Ku80, which are the key factors in NHEJ pathway, to investigate the effect of Ku silencing on the HDR efficiency in pig fetal fibroblasts. Down-regulation of Ku70 and Ku80 resulted in the promotion of the frequencies of multiple HDR pathways, including homologous recombination, single strand annealing, and single-stranded oligonucleotide-mediated DNA repair. We further evaluated the effects of Ku70 and Ku80 silencing on promoting HR-mediated knock-in efficiency in two porcine endogenous genes and found a significant increase in the amount of knock-in cells in Ku-silenced fibroblasts compared with control. The RNA interference strategy will benefit the generation of cell lines and organisms with precise genetic modifications. |
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Suppressing Ku70/Ku80 expression elevates homology-directed repair efficiency in primary fibroblasts |
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