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Problem formulation

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cristiano di pietro

on 19 January 2015

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Transcript of Problem formulation

Problem formulation
• How was it discovered?
• Why is it useful?
• How does it work?
• Examples of uses
• Future applications and prospects
• Strategies
Discovery
1990 - Richard Jorgensen's observation of the phenomena of plants which changed their colors. This process was called co-suppression.
In 2006, Drs. Fire and Mello won a Nobel Prize after reporing in "
Nature
" that the injection of double stranded RNA into C. elegans silenced the corresponding genes with complementary sequences.
Why is it useful in bioengineering?
CRISPR presents the possibility of having a cheap and highly specific tool, eligible for many applications in different fields
Future of CRISPR and applications
How does CRISPR work
- Plants and Bacteria
- Type I II and III

CRISPRi/dCas9
CasOT
CAS proteins and their functions
Cas 1 :

Cas 2 :

Cas 3/5/
6/9:
recognition of new foreign DNA
inclusion of new foreign DNA in the CRISPR sequence
Cutting of crRNAs
Cleavage of foreign DNA already in the CRISPR
Chris, Julia, Klaidas, Daniel
P1 Group 5
Examples of uses
Medicine
Bio-production
Medicine
Mice liver disease (type I tyrosinemia)
Cystic fybrosis in intestinal stem cells
Bio-production
Isopentenol production using modified E. Coli strains
Increasing crop yields, making bread wheat resistant to Powdery Mildew
Protein production using CHO cells
Biofuel production
Chinese Hamster Ovary cells
Main idea of CRISPR use for biofuel production
Pathway of bioproduction : from E. Coli to biofuel
Results of experiment : production of isopentenol
Plant genome editing using the CRISPR/Cas system
Outline of using CRISPR/Cas9 to genetically manipulate stem cells
Curing mice liver from a genetic disease
TALENs machanism as an alternative and its specifications
CRISPR/Cas9 system can be fully adapted for engeneering plant's genome
Genome editing assays in plants and its example
Explanation of Tyrosinemia (HT1)
CRISPR for in-vivo gene editing
Why the liver?
Results of the trial
Conclusion

Powdery mildew
Foilar disease
Wheat, potatoes, canola
In bread wheat it affects the flour quality and grain protein content
MLO genes
Single base pair insertions
- highest frequency
CRISPy tool
- a tool for finding off targets in CHO genomes
Discussion
-successful
-CRISPR Cas9 system + CRISPy
Results of CHO cells
RNA-Guided CRISPR Cas9 shows targeted endonuclease activity in CHO
High indel frequency obtained by all four COSMC-targeting sgRNAs
Homozygous knockout of FUT8 in CHO cells generated by CRISPR

SWOT analysis
- gRNA
- Spacers and repeats
- Pros and Cons

- Mammalians
CRISPR naturally found in bacteria
Conclusion
Future of CRISPR


Biotechnology and medicine revolution
New possibilities
Optimising CRISPR/Cas system to target the CFTR locus
Trypsinization to obtain stem cell suspension
Transfection with plasmids
Forskolin assay
Visualization of the process
Biological research
Medical research
Primary choice in bio pharmaceuticals
Big amounts of data collected
Plasmid construction and cassette desing
Full transcript