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'''Synthetic genetic array analysis''' ('''SGA''') is a [[high-throughput screening|high-throughput]] technique for exploring [[synthetic lethality|synthetic lethal]] and synthetic sick [[genetic interactions]] ([[Synthetic lethality|SSL]]).<ref name="H. Tong 2001">{{Cite journal
'''Synthetic Genetic Array analysis (SGA)''' is a [[high-throughput]] technique for exploring [[synthetic lethality|synthetic lethal]] and synthetic sick [[genetic interactions]] ([[Synthetic lethality|SSL]]) <ref name="H. Tong 2001">A. H. Tong et al., Science 294, 2364 (2001)16.</ref>. SGA allows for the systematic construction of double mutants using a combination of [[Recombinant DNA|recombinant genetic techniques]], mating and selection steps. Using SGA methodology a query gene deletion mutant can be crossed to an entire genome deletion set to identify any [[synthetic lethality|SSL]] interactions, yielding functional information of the query gene and the genes it interacts with. A large-scale application of SGA in which ~130 query genes were crossed to the set of ~5000 viable deletion mutants in yeast revealed a genetic network containing ~1000 genes and ~4000 SSL interactions <ref> A. H. Tong et al., Global Mapping of the Yeast Genetic Interaction Network, Science 303, 808 (2004) </ref>. The results of this study showed that genes with similar function tend to interact with one another and genes with similar patterns of genetic interactions often encode products that tend to work in the same pathway or complex. Synthetic Genetic Array analysis was initially developed using the model organism ''[[S.cerevisiae]]''. Methodology has since been developed to allow SGA analysis in ''[[Schizosaccharomyces pombe|S.pombe]]'' <ref>Roguev, A., Wiren, M., Weissman, J. S. & Krogan, N. J. High-throughput genetic interaction mapping in the fission yeast Schizosaccharomyces pombe. Nat Methods 4, 861-866 (2007) </ref><ref> S. J. Dixon et al., Significant conservation of synthetic lethal genetic interaction networks between distantly related eukaryotes. Proc Natl Acad Sci U S A. 105:16653-8. (2008)</ref> and ''E.coli'' <ref> Typas, A. et al. High-throughput, quantitative analyses of genetic interactions in E. coli. Nat Methods (2008). </ref>, <ref>Butland, G. et al. eSGA: E. coli synthetic genetic array analysis. Nat Methods (2008) </ref>.
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| title = Systematic Genetic Analysis with Ordered Arrays of Yeast Deletion Mutants
| doi = 10.1126/science.1065810
| journal = Science
| volume = 294
| issue = 5550
| pages = 2364–2368
| year = 2001
| pmid = 11743205
| pmc =
| bibcode = 2001Sci...294.2364T
| s2cid = 6505287
}}</ref> SGA allows for the systematic construction of double mutants using a combination of [[Recombinant DNA|recombinant genetic techniques]], mating and selection steps. Using SGA methodology a query gene deletion mutant can be crossed to an entire genome deletion set to identify any [[synthetic lethality|SSL]] interactions, yielding functional information of the query gene and the genes it interacts with. A large-scale application of SGA in which ~130 query genes were crossed to the set of ~5000 viable deletion mutants in yeast revealed a genetic network containing ~1000 genes and ~4000 SSL interactions.<ref>{{Cite journal
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| title = Global Mapping of the Yeast Genetic Interaction Network
| doi = 10.1126/science.1091317
| journal = Science
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}}</ref> The results of this study showed that genes with similar function tend to interact with one another and genes with similar patterns of genetic interactions often encode products that tend to work in the same pathway or complex. Synthetic Genetic Array analysis was initially developed using the model organism ''[[S. cerevisiae]]''. This method has since been extended to cover 30% of the ''S. cerevisiae'' genome.<ref>{{Cite journal
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}}</ref> Methodology has since been developed to allow SGA analysis in ''[[Schizosaccharomyces pombe|S.pombe]]''<ref>{{Cite journal
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| title = High-throughput genetic interaction mapping in the fission yeast Schizosaccharomyces pombe
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}}</ref> and ''E. coli''.<ref>{{Cite journal
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| title = ESGA: E. Coli synthetic genetic array analysis
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[[Image:Yeast colonies array 1536 format.jpg|thumb|300px|Arrayed yeast showing synthetic lethal interactions. Synthetic lethal interactions are those pairs of colonies with reduced or no growth.]]
 
==Background==
Synthetic Geneticgenetic Arrayarray analysis was initially developed by Tong et al. <ref name="H. Tong 2001"/> in 2001 and has since been used by many groups working in a wide range of biomedical fields. SGA utilizes the entire genome yeast knock-out set created by the yeast genome deletion project .<ref>{{cite web|url=http://sequence-www-sequence.stanford.edu/group /yeast_deletion_project/deletions3.html|title=''Saccharomyces'' Genome Deletion Project}}</ref>.
 
==Procedure==
==Synthetic genetic array analysis general procedure==
Synthetic Geneticgenetic Arrayarray analysis is generally conducted using colony arrays on petriplates at standard densities (96, 384, 768, 1536). To perform a SGA analysis in ''S.cerevisaecerevisiae'', the query gene deletion is crossed systematically with a deletion mutant array (DMA) containing every viable knockout [[Open reading frame|ORF]] of the yeast genome (currently 4786 strains).<ref>{{cite web|title=Yeast Knockout Strains
|url=http://www.openbiosystems.com/GeneExpression/Yeast/YKO/|archiveurl=https://web.archive.org/web/20111119012937/http://www.openbiosystems.com/GeneExpression/Yeast/YKO/|archivedate=November 19, 2011 |work=Open Biosystems}}</ref>. The resulting [[diploids]] are then sporulated by transferring to a media containing reduced nitrogen. The [[haploid]] progeny are then put through a series of selection platings and incubations to select for double mutants. The double mutants are screened for SSL interactions visually or using imaging software by assessing the size of the resulting colonies.
 
[[Image:Pinning robot.jpg|thumb|280px|right|Replicating yeast colonies during SGA analysis using a pinning robot]]
 
==Robotics==
Due to the large number of precise replication steps in SGA analysis, robots are widely used to perform the colony manipulations. There are a few systems specifically designed for SGA analysis, which greatly decrease the time to analyse a query gene. Generally these have a series of pins which are used to transfer cells to and from plates, with one system utilizing disposable pads of pins to eliminate washing cycles. Computer programs can be used to analyze the colony sizes from images of the plates thus automating the SGA scoring and chemical-genetics profiling.
 
== Steps for a yeast high content genome-wide genetic screening system (SGA-road map) ==
There are six major components:
 
# Mutant collection
# Material and tools for handling the mutants
# Image analysis system
# Automatic quantification and scoring system
# Confirmation approaches
# Data analysis tools
 
==See also==
*[[tetradTetrad (genetics)]]
*[[Two hybrid screening|Yeast two-hybrid]]
*[[Synthetic lethality]]
*[[Synthetic viability]]
 
==References==
Retrieved from "https://en.wikipedia.org/wiki/Synthetic_genetic_array"