Questions tagged [synthetic-biology]

Synthetic biology is an approach to engineer biology. Core to the discipline is genetic engineering, which is made more akin to engineering by adding the levels of standardization, automation, and abstraction.

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51
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3answers
2k views

Can scientists create totally synthetic life?

This particular question has been of a great deal of interest to me, especially since it dives at the heart of abiogenesis.
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What does it mean to "write an image and GIF into the DNA of bacteria"?

BBC News recently published an article saying that: An image and short film has been encoded in DNA, using the units of inheritance as a medium for storing information ... The team sequenced the ...
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1answer
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What is the criticality of the ribosome binding site relative to the start codon in prokaryotic translation?

In prokaryotic translation, how critical for efficient translation is the location of the ribosome binding site, relative to the start codon? Ideally, it is supposed to be -7b away from the start. ...
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Difference between genetic engineering and synthetic biology

I've recently seen the term synthetic biology being used to describe research involving genetic modification of organisms. What is the difference between synthetic biology and genetic engineering? Is ...
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5answers
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Introductory literature for synthetic / systems biology

I am a computer engineer (MSc in Computer Engineering) who's looking to switch into the field of synthetic / systems biology. I have a comprehensive layman's understanding of evolution, genetics, ...
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Can spider silk be synthesized to be used for 3D printing?

With the recent rise in 3D printing, I keep hearing about more novel ways to use the technology: cell 3D printing, liquid aluminum or plastic 3D printing. For example, this Ted talk deals with ...
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2answers
273 views

Is there a favoured mathematical model of bacterial growth?

One of the most regularly measured phenotypes in bacterial synthetic biology is growth. Anyone who does wet lab microbiology or synthetic biology will have performed a growth curve measurement. ...
11
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2answers
269 views

Can I use multiple bicistronic RBS sequences in a synthetic biological circuit?

The bicistroninc RBS sequences (BCDs) developed by Mutalik et al. [1] aim to remove context sensitivity from translation and therefore ensure more predictable gene expression. However, I have been ...
11
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3answers
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DNA synthesis companies: cost per base, turn-around time, codon-optimization algorithms

I would like to synthesize a 3.4kb gene (originally isolated from soil bacteria) and transform it into E. coli. There are several companies (DNA2.0, GeneScript, BlueHeron) which will synthesize the ...
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What is the highest competency possible for E coli?

I am looking to find a highly competent E coli strain. I am making a library of a ~6.6kb plasmid and I am not getting high enough efficiency. Does anyone have a suggestion of a strain/protocol with ...
9
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3answers
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Assembling small DNA parts using Golden Gate

Background I've always been told that DNA assembly can be tricky when using very small DNA parts due to low efficiency. I've also seen this when using 3A biobrick assembly to assemble promoters and ...
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143 views

How would you use readthrough as part of a genetic circuit design?

Traditionally, in synthetic biology, researchers tried to avoid some transcription phenomena (like roadblocking of tandem promoters or readthrough of weak terminators) since they are not in line with ...
9
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1answer
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How realistic is to use DNA for long term storage?

This is mainly a followup question to the recent paper Next-Generation Digital Information Storage in DNA. Personally, while I agree about the data density of the format, I can't help point out the ...
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1answer
253 views

Why is electroporation not the prefered choice transforming for Mammalian Cells (HEK)?

I see a lot of folks using different techniques for transforming mammalian cell (specifically HEK) instead of doing electroporation like I see with E.Coli (bacteria). Is there a reason for this ?
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What's the state of the art in designing and creating your own life forms?

What's the state of the art for solving the following problem. You imagine a set of features that you want a life-form to have. Just to take an arbitrary example, say you want your life-form to have a)...
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4answers
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Measuring luminescence in a fluorescence plate reader

We're considering organizing some interlaboratory work on calibrating luminescence reporters (e.g., luciferase), and one of the key questions I don't know the answer to is whether most plate readers ...
7
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3answers
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Is there any problem with using a single stop codon with a single CDS in prokaryotes?

All protein coding sequences in the iGEM Registry are supposed to end with a double stop codon. Presumably, this is to decrease the potential for read-through, which could be problematic if one is ...
7
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2answers
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How can CI repressor both activate and repress $P_{RM}$ promoter found in $\lambda$ phage?

I'm reading a paper where the authors constructed a toggle switch that uses bidirectional $P_R/P_{RM}$ promoter found in the $\lambda$ phage. There are 3 binding sites - $O_{R1}$, $O_{R2}$, $O_{R3}$ - ...
7
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1answer
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Does direction relative to origin of replication matter on small plasmids?

The recent question about forward vs. reverse strand got me thinking about directionality conventions in synthetic biology. As noted in the answer to that question, if we consider only DNA in ...
7
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2answers
855 views

Has anyone tried Gibson Assembly Optimizations?

Has anyone tried the chew back-anneal in vitro DNA assembly method (known as Gibson Assembly) for difficult sequences, like GC-rich sequences? How big constructs could you efficiently assemble? Did ...
7
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1answer
413 views

How significant is RNA degradation with removal of cap/polyA's in eukaryotes, or UTR's in prokaryotes?

Question is rather self-explanatory, but segmented into two parts. I'm attempting to make use of a repression system that employs cleaving RNA at specific areas with ribozymes with the intent of ...
6
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2answers
290 views

Compatibility between spytag/spycatcher versions

Background SpyTag and SpyCatcher are peptides which can associate via a spontaneous amide bond. Because of this, they can be fused to proteins of interest as tags to cause the proteins to bind. There ...
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Can we dilute PureExpress Cell Free Mix to increase number of reactions?

Since the PureExpress Cell Free mix is so expensive, I was wondering if it might be possible to just dilute the mix to increase the number of reactions we need to use. From this image I found: It ...
6
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2answers
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Synthetic biology using existing cells

I was watching the video at this link: http://www.bbc.co.uk/news/science-environment-17436365 The speaker says that a cell is taken and its original DNA content is stripped out and replaced with ...
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How predictably will RNA polymerase not transcribe repetitive sequences?

I am working on an (expensive) synthetic construct, which happens to have many "repetitive" sequences within it that are integral to its function. Primarily, the two sequences that are worrying me are:...
6
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1answer
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How can I convert plate reader measurements of Pichia pastoris OD to cells per ml?

I am performing experiments with the yeast Pichia pastoris where it will be beneficial to calculate product yield per cell. I plan to use a plate reader to measure OD600 of my cultures and then ...
6
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2answers
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How are Genetic Circuits Modelled?

I've read a recent Nature Methods paper by Moon T.S. et al, in which a synthetic genetic circuit consisting of layered logic gates was created. For example, the paper, a circuit is modelled in Figure ...
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2answers
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How would you effect bulk DNA gene therapy for a human?

Let's imagine that we understood DNA programming and our genome very well and realized that there were some significant flaws (we die, we need sleep, etc.) And let's imagine that we understand how to ...
6
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2answers
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Balanced vs Maximized Codon Optimization

I have worked with the optimization of several viral proteins, from different viruses/families, with different results. When Menzella, 2011 was published I tried that method. I found quite the ...
6
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1answer
355 views

What is the difference between xDNA and yDNA?

Wikipedia states that: xDNA contains expanded bases, in which a benzene ring has been added, which may pair with canon bases, resulting in four possible base-pairs (8 bases:xA-T,xT-A,xC-G,xG-C, 16 ...
6
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1answer
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Designing genes with DNAWorks: Maximum nonzero score?

I'm designing a gene in DNAWorks, and I'm unable to find a set of conditions that gives an "overall score" of zero (which is what I normally insist on). People that have used DNAWorks: what's the ...
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1answer
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How large an expression boost is typical from codon optimization?

There are lots of different codon optimization approaches out there, and a lot of different qualitative reasons to want optimization (e.g., organism codon bias, GC content, avoiding secondary ...
6
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1answer
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Is it possible to synthesize chiral version of an organism (incompatible with our pathogens)?

In theory, it should be possible to synthesize chiral (mirror image) version of some organism: with all molecules replaced with their enantiomers, e.g. L-sugars in place of our D-sugars. Direct ...
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1answer
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What happens when you tag a protease with its own degradation tag?

I've been learning about the ClipXP, ClipAP, and Lon proteases. They are proteases from the AAA+ family, which seek out proteins tagged with certain peptide sequences, unfold them, and chop them up. ...
6
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1answer
146 views

Can two hammerhead ribozymes simultaneously cleave each other?

I am working on a project in which an XOR gate is being built by having two hammerhead ribozymes simultaneously cleave each other when both are expressed, but have these ribozymes cleave a target ...
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0answers
330 views

What is the most complex biological organism (or precursors) that we have been able to synthesize from raw materials?

In the Miller–Urey experiment they produced several amino acids. I'm not sure if there were other similar experiments that got further. http://www.jcvi.org/cms/press/press-releases/full-text/article/...
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1answer
239 views

"Antibiotic resistance" equivalent in archaea for selection during cloning

I'm beginning to work with halophilic archaea and I'm trying to figure out a good way to select for cells that have taken up a plasmid. Obviously, one can't use antibiotic resistance since they are ...
5
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2answers
117 views

Are there known downsides to removing UV mutation hotspots to prevent some skin cancers (Genetic sunblock)?

Khavari et al. recently demonstrated that a significant fraction of one of the major forms of skin cancer (cutaneous squamous cell carcinomas) are associated with a mutated KNSTRN gene (a protein ...
5
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1answer
221 views

Synthetic construct with multiple ORFs not expressing

I designed a synthetic construct on paper and got it synthesized from a company. The objective was to make a vector which can be used to study both transcriptional and post-transcriptional regulation ...
5
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1answer
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Which genetic oscillator should I use to generate oscillations in range of 2-20 mins?

I'm looking to phase-separate the expression of two enzymes and hence am looking for a multi-component genetic oscillator. However, repressilators and metabolators have a large period of around 45 ...
5
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1answer
111 views

How to replace intron region in a plasmid?

I'm considering working with the plasmid pRFHUE-eGFP and would like to replace the gpdA intron (which is the eGFP promoter region) with a promoter from another organism. What would be a good strategy ...
5
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1answer
181 views

What is the relation between plasmid concentration and mRNA levels?

Suppose a simple synthetic construct, consisting of a constitutive promoter and a single gene: One of the simplest ways to model GFP transcription is to use an ODE: $\frac{d [GFP_{mRNA}]}{dt} = a - ...
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1answer
1k views

Is there a practical upper limit to amount of nucleotides or genes in a transformed plasmid?

I'm currently working on a synthetic biology project which involves working with lots of different parts. I would ultimately like to integrate these genes by transforming a single plasmid. I've heard (...
5
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2answers
934 views

What are common causes of unexpected ligation products?

I digested two plasmids, one with EcoRI and AgeI and the other with EcoRI and XmaI. Digests looked as expected, so I purified the respective fragments and set up the T4 DNA ligation (AgeI and XmaI ...
5
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2answers
180 views

Creating a cell, not from another cell. Will it be possible?

If some time in the future, we can know exactly what a cell (for example simple prokaryote bacteria) contains, (I mean, exactly which molecules, the shape of them, the density of each, everything), ...
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0answers
158 views

Could an organism theoretically produce a metamaterial-like structure? [closed]

I'm curious to know if this is physically feasible because during my reading up on synthetic biology and just general research i realise that life is capable of producing some exquisitely complex ...
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3answers
175 views

Production of plant derivatives using genetic engineered micro-organisms

I saw a Thought Emporium video where spider silk was produced by genetically modifying yeast. I have also read about companies making vanillin (vanilla flavour) using this technique. I am curious to ...
4
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1answer
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How long would you expect a designed plasmid to remain in a population of E. coli after transformation?

When transforming E. coli with a genetically-modified plasmid, how quickly/often is this plasmid "lost" because of mutation, or horizontal-transfer, or "dilution" over reproduction ...
4
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2answers
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Will new proteins incorporating new amino acids trigger an immune response?

This article reported that scientists have succeeded in adding two new bases to the quartet of A, C, G and T, resulting in non-canonical amino acid. Additionally, the bacteria in which this was done ...
4
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2answers
464 views

Has synthetic biology created new plants or animals as of 2013?

I've seen a Ted.com talk where the speaker suggests that modern science can create new micro organisms, like bacteria with a pre- defined new set of traits. But has anyone create a new species of ...