Imperial/Cell-Free/Comparison

From 2007.igem.org

(Difference between revisions)
(→Cell-Free: A Comparison of ''In Vivo'' and ''In Vitro'' Systems)
Line 18: Line 18:
</html>
</html>
-
== Cell-Free: A Comparison of ''In Vivo'' and ''In Vitro'' Systems ==
+
= Cell-Free: Advantages and Disadvantages of CFS =
{| border="1"
{| border="1"
|-
|-
-
||<center>'''In-Vitro Expression Systems'''</center>||<center>'''In-Vivo Expression Systems'''</center>
+
|width=50%|<center>'''Advantages'''</center>
 +
|width=50%|<center>'''Disadvantages'''</center>
|-
|-
-
|style="background:#eeffee"|'''Non-infectious''' because of non-proliferative nature
+
|style="background:#eeffee"|System is not an organism and is not restricted by the policies imposed on genetically modified orgnanisms (GMO)
-
|style="background:#ffeeee"|Some strains may be pathogenic
+
|style="background:#ffeeee"|Short expression lifespan because of limited energy of the system even in the presence of an ATP regenerating system
|-
|-
-
|style="background:#eeffee"|'''Process is quick and simple''' requiring only preparation of cell extract and feeding solution and subsequent addition of DNA template
+
|style="background:#eeffee"|Process is quick and simple requiring only preparation of cell extract and feeding solution and subsequent addition of DNA template
-
|style="background:#ffeeee"|Process is laborious involving DNA cloning and transformation and protein expression
+
|style="background:#ffeeee"|Expensive system has no sustained metabolism to convert cheap energy (like sugars) into useable one for the gene expression machinery
|-
|-
-
|style="background:#eeffee"|'''Good control''' can be achieved easily using modified reaction conditions such as addition of accessory elements or inhibitory factors
+
|style="background:#eeffee"|No concurrent expression of existing genome, therefore your genetically engineered device is more energy efficient
-
|style="background:#ffeeee"|Less controllability because of the presence of endogenous substances and because cells do not survive extreme conditions
+
|style="background:#ffeeee"|Less characterization and experience of use in the laboratories compared to ''E. coli''
|-
|-
-
|style="background:#eeffee"|'''Both plasmid and linear DNAs''' and can be used as templates for expression
+
|style="background:#eeffee"|No DNA mutation of your genetically engineered device because there is no DNA replication
-
|style="background:#ffeeee"|Plasmid DNAs are usually used. Linear DNAs are easily degraded by endogenous nucleases
+
|style="background:#ffeeee"|
|-
|-
-
|style="background:#eeffee"|'''Protein degradation''' is minimized by adding protease inhibitors
+
|style="background:#eeffee"|No selective pressue on your genetically engineered device because the system is non-living and does not undergo natural selection
-
|style="background:#ffeeee"|Synthesized proteins may be degraded by endogenous proteases
+
|style="background:#ffeeee"|
|-
|-
-
|style="background:#eeffee"|'''Toxic proteins''' can be synthesized in large quantities
+
|style="background:#eeffee"|No self-replication of your genetically engineered device leads to a fixed amount of DNA being expressed and more control over the rate of expression
-
|style="background:#ffeeee"|Synthesis of toxic proteins may kill the cells
+
|style="background:#ffeeee"|
|-
|-
-
|style="background:#eeffee"|'''Proteins containing unnatural amino acids''' can be achieved
+
|style="background:#eeffee"|Expression system can be quality-controlled by manipulating adjustable parameters e.g. buffers are added to maintain optimum magnesium concentrations for efficient translation; protease inhibitors can be added to minimize degradation of synthesized proteins
-
|style="background:#ffeeee"|Difficult to produce proteins containing unnatural amino acids
+
|style="background:#ffeeee"|
-
|-
+
-
|style="background:#ffeeee"|Shorter lifespan since system cannot replicate
+
-
|style="background:#eeffee"|'''Longer lifespan''' since system can replicate
+
-
|-
+
-
|style="background:#ffeeee"|More expensive because of the constant need for nutrient and energy supply
+
-
|style="background:#eeffee"|'''Less expensive''' because of the ability of the system to generate energy from relatively cheap nutrient source
+
-
|-
+
-
|style="background:#ffeeee"|Less characterized, less experience of use in the laboratories
+
-
|style="background:#eeffee"|'''Better characterized''', more experience of use in the laboratories
+
|}
|}
<br clear="all">
<br clear="all">

Revision as of 19:57, 26 October 2007


Cell-Free: Advantages and Disadvantages of CFS

Advantages
Disadvantages
System is not an organism and is not restricted by the policies imposed on genetically modified orgnanisms (GMO) Short expression lifespan because of limited energy of the system even in the presence of an ATP regenerating system
Process is quick and simple requiring only preparation of cell extract and feeding solution and subsequent addition of DNA template Expensive system has no sustained metabolism to convert cheap energy (like sugars) into useable one for the gene expression machinery
No concurrent expression of existing genome, therefore your genetically engineered device is more energy efficient Less characterization and experience of use in the laboratories compared to E. coli
No DNA mutation of your genetically engineered device because there is no DNA replication
No selective pressue on your genetically engineered device because the system is non-living and does not undergo natural selection
No self-replication of your genetically engineered device leads to a fixed amount of DNA being expressed and more control over the rate of expression
Expression system can be quality-controlled by manipulating adjustable parameters e.g. buffers are added to maintain optimum magnesium concentrations for efficient translation; protease inhibitors can be added to minimize degradation of synthesized proteins



<< What is Cell-Free? | Advantages of CFS | Our Contributions >>
Retrieved from "http://2007.igem.org/wiki/index.php/Imperial/Cell-Free/Comparison"