20190603-lab meeting_冠儀 | 賴亮全實驗室

Index

00:00 1.
index 1
00:21 2.
Engineering circular RNA for potent and stable translation in eukaryotic cellsR. Alexander Wesselhoeft, Piotr S. Kowalski & Daniel G. Anderson Nature Communications 9, Article number: 2629 (2018)
00:34 3.
Backsplicing of circRNA
01:26 4.
CircRNA are more resistant to nucleases and stable
01:55 5.
Aim
02:05 6.
Self-Splicing Introns
02:08 7.
Aim
02:08 8.
CircRNA are more resistant to nucleases and stable
02:09 9.
Backsplicing of circRNA
03:51 10.
Self-Splicing Introns
05:12 11.
IRES (Internal Ribosomal Entry Site)
06:23 12.
Results
06:24 13.
IRES (Internal Ribosomal Entry Site)
07:15 14.
Results
07:18 15.
Premuted intron-exon(PIE) splicing
08:29 16.
Addition of homology arms
08:32 17.
Premuted intron-exon(PIE) splicing
08:40 18.
Addition of homology arms
08:45 19.
Premuted intron-exon(PIE) splicing
09:45 20.
Addition of homology arms
10:15 21.
Premuted intron-exon(PIE) splicing
10:20 22.
Addition of homology arms
12:13 23.
Premuted intron-exon(PIE) splicing
12:23 24.
Addition of homology arms
17:20 25.
The effect of spacers on splicing
18:04 26.
Addition of homology arms
18:06 27.
Premuted intron-exon(PIE) splicing
19:21 28.
Addition of homology arms
19:21 29.
The effect of spacers on splicing
20:03 30.
The effect of switching to the Anabaena catalytic intron
21:41 31.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
24:41 32.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
25:40 33.
IRES efficacy in varying sequence contexts
25:45 34.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
26:02 35.
IRES efficacy in varying sequence contexts
28:15 36.
IRES efficacy in varying cell
29:11 37.
IRES efficacy in varying sequence contexts
29:14 38.
IRES efficacy in varying cell
29:20 39.
HPLC purification of circRNA from splicing reactions
32:22 40.
HPLC purification of circRNA from splicing reactions
32:25 41.
HPLC purification of circRNA from splicing reactions
33:38 42.
HPLC purification of circRNA from splicing reactions
34:31 43.
Translation efficacy of circRNA compared to linear mRNA
34:33 44.
HPLC purification of circRNA from splicing reactions
35:15 45.
Translation efficacy of circRNA compared to linear mRNA
40:42 46.
Translation efficacy of circRNA compared to linear mRNA
40:52 47.
Slide 20

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00:00 1.
index 1
00:21 2.
Engineering circular RNA for potent and stable translation in eukaryotic cellsR. Alexander Wesselhoeft, Piotr S. Kowalski & Daniel G. Anderson Nature Communications 9, Article number: 2629 (2018)
00:34 3.
Backsplicing of circRNA
01:26 4.
CircRNA are more resistant to nucleases and stable
01:55 5.
Aim
02:05 6.
Self-Splicing Introns
02:08 7.
Aim
02:08 8.
CircRNA are more resistant to nucleases and stable
02:09 9.
Backsplicing of circRNA
03:51 10.
Self-Splicing Introns
05:12 11.
IRES (Internal Ribosomal Entry Site)
06:23 12.
Results
06:24 13.
IRES (Internal Ribosomal Entry Site)
07:15 14.
Results
07:18 15.
Premuted intron-exon(PIE) splicing
08:29 16.
Addition of homology arms
08:32 17.
Premuted intron-exon(PIE) splicing
08:40 18.
Addition of homology arms
08:45 19.
Premuted intron-exon(PIE) splicing
09:45 20.
Addition of homology arms
10:15 21.
Premuted intron-exon(PIE) splicing
10:20 22.
Addition of homology arms
12:13 23.
Premuted intron-exon(PIE) splicing
12:23 24.
Addition of homology arms
17:20 25.
The effect of spacers on splicing
18:04 26.
Addition of homology arms
18:06 27.
Premuted intron-exon(PIE) splicing
19:21 28.
Addition of homology arms
19:21 29.
The effect of spacers on splicing
20:03 30.
The effect of switching to the Anabaena catalytic intron
21:41 31.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
24:41 32.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
25:40 33.
IRES efficacy in varying sequence contexts
25:45 34.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
26:02 35.
IRES efficacy in varying sequence contexts
28:15 36.
IRES efficacy in varying cell
29:11 37.
IRES efficacy in varying sequence contexts
29:14 38.
IRES efficacy in varying cell
29:20 39.
HPLC purification of circRNA from splicing reactions
32:22 40.
HPLC purification of circRNA from splicing reactions
32:25 41.
HPLC purification of circRNA from splicing reactions
33:38 42.
HPLC purification of circRNA from splicing reactions
34:31 43.
Translation efficacy of circRNA compared to linear mRNA
34:33 44.
HPLC purification of circRNA from splicing reactions
35:15 45.
Translation efficacy of circRNA compared to linear mRNA
40:42 46.
Translation efficacy of circRNA compared to linear mRNA
40:52 47.
Slide 20

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20190603-lab meeting_冠儀

Duration: 42:51, Browse: 881, Last Updated: 2019-06-13

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Q1.
What did authors do to increase the efficiency of circularization and protein translation?

00:00 1.
index 1
00:21 2.
Engineering circular RNA for potent and stable translation in eukaryotic cellsR. Alexander Wesselhoeft, Piotr S. Kowalski & Daniel G. Anderson Nature Communications 9, Article number: 2629 (2018)
00:34 3.
Backsplicing of circRNA
01:26 4.
CircRNA are more resistant to nucleases and stable
01:55 5.
Aim
02:05 6.
Self-Splicing Introns
02:08 7.
Aim
02:08 8.
CircRNA are more resistant to nucleases and stable
02:09 9.
Backsplicing of circRNA
03:51 10.
Self-Splicing Introns
05:12 11.
IRES (Internal Ribosomal Entry Site)
06:23 12.
Results
06:24 13.
IRES (Internal Ribosomal Entry Site)
07:15 14.
Results
07:18 15.
Premuted intron-exon(PIE) splicing
08:29 16.
Addition of homology arms
08:32 17.
Premuted intron-exon(PIE) splicing
08:40 18.
Addition of homology arms
08:45 19.
Premuted intron-exon(PIE) splicing
09:45 20.
Addition of homology arms
10:15 21.
Premuted intron-exon(PIE) splicing
10:20 22.
Addition of homology arms
12:13 23.
Premuted intron-exon(PIE) splicing
12:23 24.
Addition of homology arms
17:20 25.
The effect of spacers on splicing
18:04 26.
Addition of homology arms
18:06 27.
Premuted intron-exon(PIE) splicing
19:21 28.
Addition of homology arms
19:21 29.
The effect of spacers on splicing
20:03 30.
The effect of switching to the Anabaena catalytic intron
21:41 31.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
24:41 32.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
25:40 33.
IRES efficacy in varying sequence contexts
25:45 34.
Evaluation of circularization efficacy and translation from de-novo engineered precursor RNA.
26:02 35.
IRES efficacy in varying sequence contexts
28:15 36.
IRES efficacy in varying cell
29:11 37.
IRES efficacy in varying sequence contexts
29:14 38.
IRES efficacy in varying cell
29:20 39.
HPLC purification of circRNA from splicing reactions
32:22 40.
HPLC purification of circRNA from splicing reactions
32:25 41.
HPLC purification of circRNA from splicing reactions
33:38 42.
HPLC purification of circRNA from splicing reactions
34:31 43.
Translation efficacy of circRNA compared to linear mRNA
34:33 44.
HPLC purification of circRNA from splicing reactions
35:15 45.
Translation efficacy of circRNA compared to linear mRNA
40:42 46.
Translation efficacy of circRNA compared to linear mRNA
40:52 47.
Slide 20

Location

Folder name

2019

Author

鄭捷登

Branch

賴亮全教授

Created

2019-06-03 17:02:52

Last Updated

2019-06-13 12:33:54

Duration

42:51