slide1
Download
Skip this Video
Download Presentation
Potential entry points for loss of function analysis

Loading in 2 Seconds...

play fullscreen
1 / 46

Potential entry points for loss of function analysis - PowerPoint PPT Presentation


  • 387 Views
  • Uploaded on

Potential entry points for loss of function analysis. Potential entry points for loss of function analysis. Potential entry points for loss of function analysis. Dicer inactivation-in c elegans. Dcr1+/- X Dcr1+/-. Dcr-/-. Dicer inactivation-in c elegans. Dcr1+/- X Dcr1+/-. Dcr-/-. WT.

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about 'Potential entry points for loss of function analysis' - DoraAna


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
slide4

Dicer inactivation-in c elegans

Dcr1+/- X Dcr1+/-

Dcr-/-

slide5

Dicer inactivation-in c elegans

Dcr1+/- X Dcr1+/-

Dcr-/-

WT

Dcr-/-

(Science, 293, 2269)

slide6

Dicer inactivation-in c elegans

Dcr1+/- X Dcr1+/-

Dcr-/-

Dcr-/-

WT

(G & D, 15, 2654)

slide7

Dicer inactivation-in c elegans

  • Dcr1 is required for germ line formation
  • Dcr1 is not essential for later development
  • Behaviors?
  • Amplification? (maternal species)
slide8

Dicer inactivation-in c elegans

  • Dcr1 is required for germ line formation
  • Dcr1 is not essential for later development
  • Behaviors?
  • Amplification? (maternal miRNAs)

Lin4 and Let7 phenotype

slide9

Dcr1+/- X Dcr1+/-

Dcr-/-

Dicer inactivation-in zebra fish

slide10

Dcr1+/- X Dcr1+/-

Dcr-/-

Dicer inactivation-in zebra fish

Dcr1 not required for early gastralation/specifications events,

but needed for later events.

miRNA production?

slide11

Dicer inactivation-in zebra fish

(dead end: germ line depletion)

(Mir430)

(Science, 308, 833)

slide17

Dicer inactivation-in zebra fish

Dicer inactivation-in zebra fish

  • miRNAs:
  • “mostly” dispensable for cell type specification, but critical for morphogenesis--intercellular communication.
  • Probably not required for activation of cell fate specification genes

Zebrafish and c. elegans:

Differential effect on germ line formation, or non-autonomous effect?

slide18

Dicer inactivation-in zebra fish

Piwi RNA in germ line niche

Dicer function for mouse germ line formation:

WT Dicer-/-

Spindle and DNA

(G &D: 61,644)

slide19

Dicer inactivation-in zebra fish

Piwi RNA in germ line niche

Dicer function for mouse germ line formation:

(G &D: 61,644)

slide21

Dicer inactivation-in mouse

T expression Oct 4 expression

(Nature Genetics, 35, 215)

slide22

Dicer inactivation-in mouse

T expression Oct 4 expression

What cells are missing?

(Nature Genetics, 35, 215)

slide23

Dicer inactivation-in mouse

Conditional alleles:

Yeast,

C elegans,

Drosophila,

Zebrafish,

Mammals,

slide24

Dicer inactivation-in mouse

Conditional alleles:

Yeast,

C elegans,

Drosophila,

Zebrafish,

Mammals: integrases Cre/LoxP, FRT/Flpe, PhiC31

PLoS ONE. 2: e162

slide25

Dicer inactivation-in mouse

Conditional alleles:

(G &D: 19, 489)

slide26

Dicer inactivation-in mouse

What do you expect?

(G &D: 19, 489)

slide27

Dicer inactivation-in mouse

What do you expect?

(G &D: 19, 489)

slide28

Dicer inactivation-in mouse

miRNAs: required for differentiation

(G &D: 19, 489)

slide30

Functional comparison between TFs and miRNA

miRNA vs transcription factors

Mode of

action post-transcription transcription

slide31

Dicer inactivation-in mouse

Heterochromatin formation affected in Dicer-/- ES cells

?

Methylation pattern

miRNA

?

Dnmts

(G &D: 19, 489)

slide32

Dicer inactivation-in mouse

Heterochromatin formation affected in Dicer-/- ES cells

?

Methylation pattern

miRNA

?

Dnmts

(G &D: 19, 489)

slide33

Dicer inactivation-in mouse

Specific functions:

  • 110, 000 miRNA molecules/ES cell.
  • More than half belongs to four clusters, related with cell cycle
  • more than 300 different types

(PNAS: 104, 18097)

slide34

Dicer inactivation-in mouse

Zp3- Cre to produce oocyte-specific deletion

slide35

Dicer inactivation-in mouse

MyoD-Cre for muscle

(DB: 311, 359)

slide40

Deletion of individual miRNAs

Only two examples so far in mouse: Mir208 and Mir155

slide41

Deletion of individual miRNAs

Only two examples so far in mouse: Mir208 and Mir155

Accompanied by abnormal immune cell deposition

+/+ -/-

Science: 316, 604 and 608)

slide42

Deletion of individual miRNAs

Only two examples so far in mouse: Mir208 and Mir155

Science: 316, 604 and 608)

slide43

Deletion of individual miRNAs

Only two examples so far in mouse: Mir208 and Mir155

Survival:

Pathogen challenge

Science: 316, 604 and 608)

slide44

Deletion of individual miRNAs

Only two examples so far in mouse: Mir208 and Mir155

IG production:

Pathogen challenge

Science: 316, 604 and 608)

slide45

Deletion of individual miRNAs

Only two examples so far in mouse: Mir208 and Mir155

T-cell proliferation

Science: 316, 604 and 608)

slide46

Redundancy

or the functional mode of miRNA?

ad