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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Adh2 promoter in saccharomyces cerevisiae

I'm looking at expressing a protein in saccharomyces cerevisiae using the Adh2 promoter. My understanding is that the gene will be repressed by the presence of glucose, but when glucose runs out it ...
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How would it be possible to engineer cows to have more offspring?

Cows usually only give birth to one calf per year and only several in their lifetime. What would we have to genetically alter so they can have more offspring, more frequently. I read that their is a ...
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DNA Design for Multi-Site Restriction Enzymes

I have to use the SacII restriction enzyme, which requires multiple recognition sites to efficiently cut DNA, for a DNA assembly. From my understanding, multiple copies of DNA holding only a single ...
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For a biological AND Gate to work, do the two inputs need to arrive at the same time?

I understand that AND gates require the two inputs to be present for the output to be produced. However, if one input arrives faster than the other input does the output still get produced? For ...
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What is the structure of PduK in the Pdu BMC?

My lab is creating 1,2-propanediol utilization bacterial microcompartments (Pdu BMCs) with an operon containing genes for PduA, PduB, PduJ, PduK, PduN, PduU, and PduT. According to Mayer et al., all ...
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6 votes
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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 ...
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2 votes
1 answer
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Using PCR to add the overhangs to gBlocks for NEB HiFi

Does adding the overhangs to gBlocks for NEB Hifi using PCR pose a big problem? The IDT website suggests that one should design the gBlocks with the overhangs. However, if I did not do this, will it ...
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3 votes
1 answer
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T-vector creation

I'm working on TA cloning for the first time and I need to turn some existing plasmid backbones into T-vectors. I'm using a Klenow fragment (3' -> 5' exo-) to dA-tail my inserts. Is there any ...
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Identifying Promoters in Non-homologous Sequences within the Same Organism

In this paper, the authors cloned a GPD promoter for use in driving a GFP gene from Lentinula edodes using a set of provided primers. The primers were derived from the following accession GQ457137.1 ...
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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 ...
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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 ...
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Impact of Terminator Efficiency in Genetic Constructs

I was looking at the iGEM Bioregistry of Terminator parts which offer varying degrees of termination efficiency. I am wondering why studies into combinatorial synthesis of genetic circuits for ...
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What is the life of cell-free genetic circuits?

Can genetic circuits be tested in a cell-free environment? What would be their life if we keep them at room temperature? The circuits which are composed of DNA-based components are commonly referred ...
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8 votes
1 answer
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Does gene orientation 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 ...
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Forward or Reverse Strand: Is there a difference when encoding genetic devices?

Background: In synthetic biology, and also in nature, there are lots of examples of genes in both the forward and reverse orientation. It seems in synthetic biology/bioengineering, most genetic ...
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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 ...
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9 votes
2 answers
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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 ...
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4 votes
2 answers
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What genes are required to make E. coli photosynthetic?

"All" of the genes for bacterial photosynthesis were discovered in a gene cluster almost 40 years ago. Marrs, J.Bact. What more is needed to make E. coli photosynthetic?
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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 ...
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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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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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12 votes
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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. ...
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11 votes
2 answers
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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 ...
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7 votes
4 answers
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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 ...
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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 ...
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4 votes
1 answer
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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 ...
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3 votes
1 answer
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Are 2A cleavage sites functional in bacteria?

In mammalian cell lines, 2A self-cleaving peptides are frequently used to for expression of multiple genes from a single transcript. They've got some issues, e.g., leaving some proteins as fusions, ...
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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 ...
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2 votes
2 answers
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Why is the start of my coding sequence ATG and not TAC?

I am engineering a set of genetic sequences and have come across a surprisingly basic point of confusion that seems to have fallen through the cracks regarding coding sequences. The standard start ...
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2 votes
1 answer
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Is synthesizing DNA in-house feasible?

I've been reading up about DNA writing, and it seems there are machines available around the five-figure US dollar mark for DNA synthesis, and I'm considering picking one up for my lab. We normally ...
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-1 votes
2 answers
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What is the energy required for storing and performing computations in synthetic computational biological cells?

I want to know the energy costs of various computations performed through synthetic biological circuits in terms of ATP. I am primarily interested in storing of bits, addition, and multiplication ...
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How can I get started in molecular biology? [closed]

I have a background in computer science and math and am interested in learning molecular / synthetic biology - what resources do people recommend?
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How to estimate mRNA counts from Relative Promoter Units (RPU) or RNAP per second (PoPS) in E. coli?

I need a rough estimate on how to translate RPU or Polymerases Per Second (PoPS) to mRNA count (or even better protein count) in E. coli. I understand that any number we come up with will be a very ...
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2 votes
1 answer
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Source for 'EVERY living organism uses ATP as energy carrier', and can we make a synthetic one that doesn't?

I have read it repeatedly that 'all living organisms use ATP as an energy currency etc' and 'some use GTP in addition'. However I am yet to find a reliable resource, rather than just quora/reddit/...
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4 votes
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Replacing, instead of repairing, DNA

I've been doing some light reading on DNA damage theory of aging. One of the main ideas from that theory that I got is that the accumulation of damage in our DNA is one of the biggest causes of why we ...
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1 answer
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What are low-accumulating genotypes? [closed]

What are low-accumulating genotypes? And how does it differ from high-accumulating genotypes?
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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 ...
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What is the purpose of the AraC gene in pGLO?

The AraC gene activates in pGLO from my understanding in the presence of arabinose. Arabinose is present as a metabolite from other metabolic in the e.coli cell. Is it that there's no termination ...
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1 vote
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Papers where protein oligomerization has been engineered away

I have a protein that acts as a monomer, but appears to have a relatively high self-affinity in vivo (measured using transient transfection of a plasmid coding for the protein), which is annoying ...
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0 votes
2 answers
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A synthetic sieve organ known as kliver?

in this video at 5:28, the narrator talks about a "vat-grown all-purpose sieve organ" called Kliver that would do away with both liver and kidney transplant. But i don't seem to find online resources ...
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MIT's Programming Language for Cells

I have read a lot about the programming language, Cello, that MIT created to program DNA; however, I cannot seem to find any sources online that shows you how to program with Cello. Are there any good ...
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1 vote
1 answer
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Could the protein dystrophin be artificially synthesised?

Could the protein dystrophin be artificially synthesised and if so could patients with DMD (Duchenne Muscular Dystrophy) benefit from it? //Now I don't have much scientific background other than a ...
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Dealing with Repetitive Bases in DNA?

I have been given a project involving a plasmid that contains long stretches of Adenine (60 or 120 bases each). These PolyA stretches are interrupted by the occasional G or C. I understand that ...
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Assay for Glycosyltransferase

I want to do an assay to see, if an ordered enzyme is active or not. It's a glucosyltransferase from S. mutans which hydrolyses sucrose into fructose and glucose. It then connects the glucose-monomers ...
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37 votes
3 answers
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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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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. ...
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Storage of bacteria

Why are strains of bacteria stored when they are inactive (frozen)? What is the problem with storing growing cultures for long periods?
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3 votes
1 answer
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What's the difference between prions and prion-like proteins?

If I added a prion domain to a protein, does that make the protein a prion-like protein or would it be considered a prion at that point? I'm trying to understand what prions are, how they aggregate ...
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Is cell-free Life possible? [closed]

I think most of the people on this site know about the famous Baltimore paper describing the ability of the poliovirus to replicate itself without the need of cells but I want to expand on the issue ...
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