2013 TzuChiU_Formosa Team Team Membe Members rs Hypnoseq. - - PowerPoint PPT Presentation

▶
2013 tzuchiu formosa team team
SMART_READER_LITE
LIVE PREVIEW

2013 TzuChiU_Formosa Team Team Membe Members rs Hypnoseq. - - PowerPoint PPT Presentation

Dec 23, 2023 27 likes •465 views

2013 TzuChiU_Formosa Team Team Membe Members rs Hypnoseq. Hypnos Sequence Right before Us Right before Us Infections caused by resistant microorganisms often fail to respond to conventional treatment, resulting in prolonged illness,

slide-1
SLIDE 1

2013 TzuChiU_Formosa

slide-2
SLIDE 2

Team Team Membe Members rs

slide-3
SLIDE 3

Hypnos Sequence

Hypnoseq.

slide-4
SLIDE 4

Right before Us

slide-5
SLIDE 5
  • Infections caused by resistant microorganisms often fail

to respond to conventional treatment, resulting in prolonged illness, greater risk of death and higher costs.

  • New resistance mechanisms have emerged, making the

latest generation of antibiotics virtually ineffective. －World Health Organization

Right before Us

slide-6
SLIDE 6
slide-7
SLIDE 7

1

Introduction

We We ha have ve fea eatur tures es for r ev ever ery ste y step of p of th the pro e proje ject. ct.

slide-8
SLIDE 8
slide-9
SLIDE 9

chloramphenicol

Designed four parts of Cam-antisenses － specific

slide-10
SLIDE 10

Pri Primer Des mer Design ign

Obtain antibiotic resistance sequence Random selection to seek high specificity regions Reverse the above sequence Add the restriction enzyme site at the end of primers

slide-11
SLIDE 11

Hypno Hypnoseq. seq.

How doe How does it w s it work?

  • rk?

RISC RISC?

Dice Dicer? r?

in Proka in Prokaryotes ryotes?

?

" Are organisms able to perform an easier mechanism to carry out our desired plan? "

slide-12
SLIDE 12

2

System Design

1

Introduction

We We ha have ve fea eatur tures es for r ev ever ery ste y step of p of th the pro e proje ject. ct.

slide-13
SLIDE 13

13

IPTG

Target mRNA

slide-14
SLIDE 14

2

System Design

1

Introduction

3

Results

We We ha have ve fea eatur tures es for r ev ever ery ste y step of p of th the pro e proje ject. ct.

slide-15
SLIDE 15

Gr Group 1

  • up 1

pSB1C3 vector antisense

Colony PCR

slide-16
SLIDE 16

Lac I PCR

from pET11d (E.c .coli)

slide-17
SLIDE 17

 pSB1C3 group Clone Lac I Construct the system Transformation Quantify the system

In P In Prog rogress ress

slide-18
SLIDE 18

Gr Group 2

  • up 2

pET11d vector antisense

slide-19
SLIDE 19

Amp

Colony

pET11d

slide-20
SLIDE 20

2

System Design

4

Future & Application

1

Introduction

3

Results

We We ha have ve fea eatur tures es for r ev ever ery ste y step of p of th the pro e proje ject. ct.

slide-21
SLIDE 21

IPTG

add

Cam

add

Plating Plating

slide-22
SLIDE 22

Decrease

the percentage of

Antibiotic Resistance

  • r
slide-23
SLIDE 23

Applications Applications

  • Health/Medicine：Knockdown pathogenic genes of bacteria
  • New Application ：on-off switch
slide-24
SLIDE 24

CRISPR

(Clustered Regularly Interspaced Short Palindromic Repeats)

slide-25
SLIDE 25

Brand New Design ! CRISPR/Cas9 system

slide-26
SLIDE 26

Conjugation CRISPR/Cas9

OriTR J01003
slide-27
SLIDE 27

2

System Design

4

Future & Application

1

Introduction

3

Results

5

Achieve- ment

We We ha have ve fea eatur tures es for r ev ever ery ste y step of p of th the pro e proje ject. ct.

slide-28
SLIDE 28

28

Ampicillin antisense 1(BBa_K1222997) Ampicillin antisense 2(BBa_K1222996) Ampicillin antisense 3(BBa_K1222995) Ampicillin antisense 4(BBa_K1222994) GFP antisense 1(BBa_K1222998) GFP antisense 2(BBa_K1222989) GFP antisense 3(BBa_K1222988) Lac operator+CamR antisense 1(BBa_K1222984) Lac operator+CamR antisense 2(BBa_K1222983) Lac operator+CamR antisense 3(BBa_K1222982) Lac operator+CamR antisense 4(BBa_K1222981) Lac operator+GFP antisense 1(BBa_K1222987) Lac operator+GFP antisense 2(BBa_K1222986) Lac operator+GFP antisense 3(BBa_K1222985) Lac promoter+LacI(BBa_K1222002) T7 promoter+lac operator+antisense +T7 terminator (BBa_K1222004, BBa_K1222005) T7 promoter+lac operator+antisense+T7 terminator+Lac promoter+LacI(BBa_K1222000, BBa_K1222001)

Completed.

slide-29
SLIDE 29
slide-30
SLIDE 30

Human Human Prac Practices tices

  • Spe

Speech ech

  • Fa

Facebook cebook Pag Page

  • SB Work

SB Workshop shop

slide-31
SLIDE 31

Huma Human Prac n Practic tices es

slide-32
SLIDE 32

Human Human Prac Practices tices

  • Spe

Speech ech

  • Fa

Facebook cebook Pag Page

  • SB Work

SB Workshop shop

slide-33
SLIDE 33
slide-34
SLIDE 34

Human Human Prac Practices tices

  • Spe

Speech ech

  • Fa

Facebook cebook Pag Page

  • SB Work

SB Workshop shop

slide-35
SLIDE 35

Acknowledgements Acknowledgements

slide-36
SLIDE 36

Shu-Jun Chiu Jui-Hung Yen Ji-Hshiung Chen Ingrid Liu Jeng-Woei Lee Chun-Yao Chen Guang-Huey Lin

Ins Instruc tructor tor Te Techni chnical cal Su Suppo pports rts

slide-37
SLIDE 37

Advi Advisor sors

Hung-Jen Chien Hsin-Jung Tien Chia-Sheng Pai Lu-Chu Ke Li-Yi Lin Kai-Chi Liang Ni-Chun Chung Yao-Hua Hew Ruo-Yu Wang Hsien-Ting Huang

slide-38
SLIDE 38
slide-39
SLIDE 39

Refer References ences

Hale, C. R., et al. (2009). "RNA-guided RNA cleavage by a CRISPR RNA-Cas protein complex." Cell 139(5): 945-956. Hsu, P. D., et al. (2013). "DNA targeting specificity of RNA-guided Cas9 nucleases." Nat Biotechnol 31(9): 827-832. Huang, L., et al. (2013). "Efficient and specific gene knockdown by small interfering RNAs produced in bacteria." Nat Biotechnol 31(4): 350-356. Wang, H., et al. (2013). "One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering." Cell 153(4): 910-918. Wiedenheft, B., et al. (2012). "RNA-guided genetic silencing systems in bacteria and archaea." Nature 482(7385): 331-338.

slide-40
SLIDE 40

Thank you Thank you for your for your attenti attention.

  • n.
slide-41
SLIDE 41
slide-42
SLIDE 42

Ampr mRNA

protein

Camr Ampr Kanr

Amp Amp protein

slide-43
SLIDE 43

Anti-sense Ampr mRNA

IPTG

Ampr mRNA

protein Amp

Camr Ampr Kanr Ampr mRNA