• 00:00 1.
    index 1
  • 00:59 2.
    Slide 2
  • 01:26 3.
    Introduction
  • 03:02 4.
    Slide 4
  • 04:19 5.
    Examination of universally expressed circRNAs
  • 04:59 6.
    Treated HeLa cells with 8 stressors followed by the examination of universally expressed circRNAs
  • 07:10 7.
    circRNAs are degraded upon Poly I:C treatment
  • 08:12 8.
    Global reduction of high confidence circRNAs upon poly(I:C) for 6 h (C) or EMCV for 24 h
  • 08:13 9.
    circRNAs are degraded upon Poly I:C treatment
  • 09:12 10.
    Treated HeLa cells with 8 stressors followed by the examination of universally expressed circRNAs
  • 10:26 11.
    circRNAs are degraded upon Poly I:C treatment
  • 11:26 12.
    Treated HeLa cells with 8 stressors followed by the examination of universally expressed circRNAs
  • 13:44 13.
    circRNAs are degraded upon Poly I:C treatment
  • 13:46 14.
    Global reduction of high confidence circRNAs upon poly(I:C) for 6 h (C) or EMCV for 24 h
  • 13:54 15.
    circRNAs are degraded upon Poly I:C treatment
  • 13:55 16.
    Global reduction of high confidence circRNAs upon poly(I:C) for 6 h (C) or EMCV for 24 h
  • 18:02 17.
    circRNAs are degraded upon Poly I:C treatment
  • 18:02 18.
    Treated HeLa cells with 8 stressors followed by the examination of universally expressed circRNAs
  • 18:02 19.
    circRNAs are degraded upon Poly I:C treatment
  • 18:04 20.
    Global reduction of high confidence circRNAs upon poly(I:C) for 6 h (C) or EMCV for 24 h
  • 18:11 21.
    Slide 9
  • 18:26 22.
    RNase L “may” be responsible for circRNA degradation upon poly(I:C) and viral stimulations
  • 21:45 23.
    Slide 9
  • 21:47 24.
    RNase L “may” be responsible for circRNA degradation upon poly(I:C) and viral stimulations
  • 22:14 25.
    Slide 11
  • 22:16 26.
    RNase L “may” be responsible for circRNA degradation upon poly(I:C) and viral stimulations
  • 22:22 27.
    Slide 11
  • 24:23 28.
    RNase L is responsible for global circRNA degradation in cells treated with poly(I:C)
  • 26:02 29.
    The endonuclease activity of RNase L is requiredfor circRNA degradation
  • 26:35 30.
    Slide 14
  • 26:36 31.
    The endonuclease activity of RNase L is requiredfor circRNA degradation
  • 28:19 32.
    Slide 14
  • 28:19 33.
    The endonuclease activity of RNase L is requiredfor circRNA degradation
  • 28:33 34.
    Slide 14
  • 28:39 35.
    The endonuclease activity of RNase L is requiredfor circRNA degradation
  • 28:41 36.
    Slide 14
  • 28:52 37.
    Slide 15
  • 29:02 38.
    In vitro binding and competition screening assays with purified immune factors
  • 29:05 39.
    Slide 15
  • 29:27 40.
    In vitro binding and competition screening assays with purified immune factors
  • 30:50 41.
    Slide 15
  • 30:54 42.
    In vitro binding and competition screening assays with purified immune factors
  • 31:43 43.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 37:21 44.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 37:28 45.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 37:28 46.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 37:29 47.
    dsRBMs of PKR are required for binding to circPOLR2A
  • 37:30 48.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 38:00 49.
    dsRBMs of PKR are required for binding to circPOLR2A
  • 39:48 50.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 39:49 51.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 39:49 52.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 40:22 53.
    dsRBMs of PKR are required for binding to circPOLR2A
  • 40:24 54.
    circRNAs prefer to bind to nucleic acid receptors with antiviral activity
  • 40:43 55.
    dsRBMs of PKR are required for binding to circPOLR2A
  • 40:44 56.
    PKR is an IFN-inducible Ser/Thr protein kinase directly activated by dsRNA
  • 41:25 57.
    Slide 21
  • 43:29 58.
    Slide 22
  • 43:40 59.
    Slide 23
  • 43:41 60.
    Slide 22
  • 43:43 61.
    Slide 23
  • 44:08 62.
    Slide 24
  • 45:11 63.
    Comparison of paired circular and linear RNA structures by SHAPE-MaP
  • 45:14 64.
    Slide 24
  • 45:38 65.
    Comparison of paired circular and linear RNA structures by SHAPE-MaP
  • 46:51 66.
    Linear POLR2A is relatively unstructured within circPOLR2A-forming region
  • 48:04 67.
    circRNAs have distinct structural conformation from their linear cognate RNAs
  • 48:35 68.
    Many examined circRNAs tend to form imperfect duplex RNA regions
  • 49:08 69.
    Slide 29
  • 49:12 70.
    Slide 30
  • 50:24 71.
    Slide 31
  • 50:52 72.
    Slide 32
  • 51:00 73.
    Slide 33
  • 51:21 74.
    Most examined circRNAs could form 1–4 dsRNA regions.Hypothesis : Per cell may contains > 10,000~20,000 intra-dsRNA regions and that such circRNAs represent a type of endogenous inhibitors for PKR.
  • 52:41 75.
    Overexpression of circRNAs in Hela cell
  • 53:20 76.
    Most examined circRNAs could form 1–4 dsRNA regions.Hypothesis : Per cell may contains > 10,000~20,000 intra-dsRNA regions and that such circRNAs represent a type of endogenous inhibitors for PKR.
  • 53:24 77.
    Overexpression of circRNAs in Hela cell
  • 53:53 78.
    Overexpression of circPOLR2A in HeLa cells inhibits PKR phosphorylation upon poly(I:C) stimulation
  • 54:51 79.
    Overexpression of the non-dsRNA-containing circRNA in HeLa cells does not suppress PKR phosphorylation upon poly(I:C) stimulation.
  • 56:17 80.
    Slide 38
  • 56:40 81.
    Slide 39
  • 56:42 82.
    Reduced PKR activation in RNase L KO HeLa cells stimulated with poly(I:C)
  • 56:42 83.
    Slide 39
  • 57:07 84.
    Reduced PKR activation in RNase L KO HeLa cells stimulated with poly(I:C)
  • 57:48 85.
    Overexpression of circPOLR2A in HeLa cells inhibitsPKR phosphorylation upon EMCV infection
  • 58:11 86.
    Slide 42
  • 58:18 87.
    Dysregulated circRNA expression and PKR activation in SLE
  • 59:47 88.
    circRNA reduction in PBMCs derived from SLE patients
  • 1:00:15 89.
    Slide 45
  • 1:02:01 90.
    Global reduction of circRNA expression in SLE patients (RNA-seq)
  • 1:02:39 91.
    Slide 47
  • 1:03:08 92.
    circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in PBMCs isolated from three SLE patients
  • 1:04:10 93.
    circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in T cells isolated from three SLE patients
  • 1:04:11 94.
    circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in PBMCs isolated from three SLE patients
  • 1:04:27 95.
    circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in T cells isolated from three SLE patients
  • 1:04:31 96.
    circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in PBMCs isolated from three SLE patients
  • 1:04:34 97.
    circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in T cells isolated from three SLE patients
  • 1:04:41 98.
    Conclusion
  • 1:04:54 99.
    Slide 51
  • 1:05:45 100.
    Highlights
  • 1:06:16 101.
    circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in PBMCs isolated from three SLE patients
  • Index
  • Notes
  • Comment
  • Fullscreen
20190520-lab meeting_捷登
Duration: 1:07:08, Browse: 616, Last Updated: 2019-06-13
Keypoint
  1. 1.
    f8bf050378a020b030249b59422e37af.png
    • 00:00 1.
      index 1
    • 00:59 2.
      Slide 2
    • 01:26 3.
      Introduction
    • 03:02 4.
      Slide 4
    • 04:19 5.
      Examination of universally expressed circRNAs
    • 04:59 6.
      Treated HeLa cells with 8 stressors followed by the examination of universally expressed circRNAs
    • 07:10 7.
      circRNAs are degraded upon Poly I:C treatment
    • 08:12 8.
      Global reduction of high confidence circRNAs upon poly(I:C) for 6 h (C) or EMCV for 24 h
    • 08:13 9.
      circRNAs are degraded upon Poly I:C treatment
    • 09:12 10.
      Treated HeLa cells with 8 stressors followed by the examination of universally expressed circRNAs
    • 10:26 11.
      circRNAs are degraded upon Poly I:C treatment
    • 11:26 12.
      Treated HeLa cells with 8 stressors followed by the examination of universally expressed circRNAs
    • 13:44 13.
      circRNAs are degraded upon Poly I:C treatment
    • 13:46 14.
      Global reduction of high confidence circRNAs upon poly(I:C) for 6 h (C) or EMCV for 24 h
    • 13:54 15.
      circRNAs are degraded upon Poly I:C treatment
    • 13:55 16.
      Global reduction of high confidence circRNAs upon poly(I:C) for 6 h (C) or EMCV for 24 h
    • 18:02 17.
      circRNAs are degraded upon Poly I:C treatment
    • 18:02 18.
      Treated HeLa cells with 8 stressors followed by the examination of universally expressed circRNAs
    • 18:02 19.
      circRNAs are degraded upon Poly I:C treatment
    • 18:04 20.
      Global reduction of high confidence circRNAs upon poly(I:C) for 6 h (C) or EMCV for 24 h
    • 18:11 21.
      Slide 9
    • 18:26 22.
      RNase L “may” be responsible for circRNA degradation upon poly(I:C) and viral stimulations
    • 21:45 23.
      Slide 9
    • 21:47 24.
      RNase L “may” be responsible for circRNA degradation upon poly(I:C) and viral stimulations
    • 22:14 25.
      Slide 11
    • 22:16 26.
      RNase L “may” be responsible for circRNA degradation upon poly(I:C) and viral stimulations
    • 22:22 27.
      Slide 11
    • 24:23 28.
      RNase L is responsible for global circRNA degradation in cells treated with poly(I:C)
    • 26:02 29.
      The endonuclease activity of RNase L is requiredfor circRNA degradation
    • 26:35 30.
      Slide 14
    • 26:36 31.
      The endonuclease activity of RNase L is requiredfor circRNA degradation
    • 28:19 32.
      Slide 14
    • 28:19 33.
      The endonuclease activity of RNase L is requiredfor circRNA degradation
    • 28:33 34.
      Slide 14
    • 28:39 35.
      The endonuclease activity of RNase L is requiredfor circRNA degradation
    • 28:41 36.
      Slide 14
    • 28:52 37.
      Slide 15
    • 29:02 38.
      In vitro binding and competition screening assays with purified immune factors
    • 29:05 39.
      Slide 15
    • 29:27 40.
      In vitro binding and competition screening assays with purified immune factors
    • 30:50 41.
      Slide 15
    • 30:54 42.
      In vitro binding and competition screening assays with purified immune factors
    • 31:43 43.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 37:21 44.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 37:28 45.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 37:28 46.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 37:29 47.
      dsRBMs of PKR are required for binding to circPOLR2A
    • 37:30 48.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 38:00 49.
      dsRBMs of PKR are required for binding to circPOLR2A
    • 39:48 50.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 39:49 51.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 39:49 52.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 40:22 53.
      dsRBMs of PKR are required for binding to circPOLR2A
    • 40:24 54.
      circRNAs prefer to bind to nucleic acid receptors with antiviral activity
    • 40:43 55.
      dsRBMs of PKR are required for binding to circPOLR2A
    • 40:44 56.
      PKR is an IFN-inducible Ser/Thr protein kinase directly activated by dsRNA
    • 41:25 57.
      Slide 21
    • 43:29 58.
      Slide 22
    • 43:40 59.
      Slide 23
    • 43:41 60.
      Slide 22
    • 43:43 61.
      Slide 23
    • 44:08 62.
      Slide 24
    • 45:11 63.
      Comparison of paired circular and linear RNA structures by SHAPE-MaP
    • 45:14 64.
      Slide 24
    • 45:38 65.
      Comparison of paired circular and linear RNA structures by SHAPE-MaP
    • 46:51 66.
      Linear POLR2A is relatively unstructured within circPOLR2A-forming region
    • 48:04 67.
      circRNAs have distinct structural conformation from their linear cognate RNAs
    • 48:35 68.
      Many examined circRNAs tend to form imperfect duplex RNA regions
    • 49:08 69.
      Slide 29
    • 49:12 70.
      Slide 30
    • 50:24 71.
      Slide 31
    • 50:52 72.
      Slide 32
    • 51:00 73.
      Slide 33
    • 51:21 74.
      Most examined circRNAs could form 1–4 dsRNA regions.Hypothesis : Per cell may contains > 10,000~20,000 intra-dsRNA regions and that such circRNAs represent a type of endogenous inhibitors for PKR.
    • 52:41 75.
      Overexpression of circRNAs in Hela cell
    • 53:20 76.
      Most examined circRNAs could form 1–4 dsRNA regions.Hypothesis : Per cell may contains > 10,000~20,000 intra-dsRNA regions and that such circRNAs represent a type of endogenous inhibitors for PKR.
    • 53:24 77.
      Overexpression of circRNAs in Hela cell
    • 53:53 78.
      Overexpression of circPOLR2A in HeLa cells inhibits PKR phosphorylation upon poly(I:C) stimulation
    • 54:51 79.
      Overexpression of the non-dsRNA-containing circRNA in HeLa cells does not suppress PKR phosphorylation upon poly(I:C) stimulation.
    • 56:17 80.
      Slide 38
    • 56:40 81.
      Slide 39
    • 56:42 82.
      Reduced PKR activation in RNase L KO HeLa cells stimulated with poly(I:C)
    • 56:42 83.
      Slide 39
    • 57:07 84.
      Reduced PKR activation in RNase L KO HeLa cells stimulated with poly(I:C)
    • 57:48 85.
      Overexpression of circPOLR2A in HeLa cells inhibitsPKR phosphorylation upon EMCV infection
    • 58:11 86.
      Slide 42
    • 58:18 87.
      Dysregulated circRNA expression and PKR activation in SLE
    • 59:47 88.
      circRNA reduction in PBMCs derived from SLE patients
    • 1:00:15 89.
      Slide 45
    • 1:02:01 90.
      Global reduction of circRNA expression in SLE patients (RNA-seq)
    • 1:02:39 91.
      Slide 47
    • 1:03:08 92.
      circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in PBMCs isolated from three SLE patients
    • 1:04:10 93.
      circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in T cells isolated from three SLE patients
    • 1:04:11 94.
      circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in PBMCs isolated from three SLE patients
    • 1:04:27 95.
      circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in T cells isolated from three SLE patients
    • 1:04:31 96.
      circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in PBMCs isolated from three SLE patients
    • 1:04:34 97.
      circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in T cells isolated from three SLE patients
    • 1:04:41 98.
      Conclusion
    • 1:04:54 99.
      Slide 51
    • 1:05:45 100.
      Highlights
    • 1:06:16 101.
      circPOLR2A overexpression attenuates aberrant PKR and EIF2α phosphorylation in PBMCs isolated from three SLE patients
    Location
    Folder name
    2019
    Author
    鄭捷登
    Branch
    賴亮全教授
    Created
    2019-05-20 10:33:55
    Last Updated
    2019-06-13 12:49:55
    Duration
    1:07:08