...of a generic promoter). A DIO opsin allows cell type specificity dependent on the promoter of the cre recombinase...neuronal specificity through promoter-directed expression alone is extremely limited. Therefore, highly ...communicated to me as one of these extremely high expressing promoters). Conduct a time course experiment...for sufficiency of your circuit of interest in promoting reward is a self-stimulation experiment wherein...developed many of these AAV constructs with various promoters and other genetic manipulations to help restrict...specific neuronal subtype. However, the length of promoter that can be used in an AAV construct is limited...highly expressing, generic neuronal promoters such as synapsin or camKIIα drive most of the constructs. As ...

...dependent promoters, Baojun Wang’s lab developed a system that can activate sigma-54 dependent promoters (Liu...activation of sigma-54 type promoters. The system was functional in activating promoters in Pseudomonas syringae...that utilizes phage recombination machinery to promote homologous recombination of linear DNA fragments...regulate gene expression were limited to inducible promoters or direct gene knockout. In contrast, CRISPR offers...systems, catalytically dead (dCas9) targeted to a promoter or gene body can repress transcription by physically...and can be more effective targeting either the promoter or the coding sequence and the template or non-template...to multiplex repression. Using a Tet responsive promoter to drive expression of a catalytically dead Cas9...

...hybrid promoter. Although the DE3/T7 RNAP system works well for most experiments, the lac promoter can “...expression from high-copy number plasmids and powerful promoters will greatly exceed that of any native host protein... accomodate high protein loads using inducible promoter systems (which will be discussed later). In addition...Kanamycin (pREP4) Cis-repression of the E. coli T5 promoter (found on vectors such as pQE or similar), inducible...above, many expression plasmids utilize inducible promoters, which are 'inactive' until an inducer such as...the culture to make protein.  The DE3 lysogen/T7 promoter combination is the most popular induction system...available to transcribe the gene of interest from a T7 promoter on the plasmid. Many commercial strains carry ...

...all chemically inducible promoters; 3) improving the strength of the promoters used; and 4) optimizing ...fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various pieces...sequences. Several pre-constructed promoter-selection marker or promoter-inducible expression related vectors...ideal promoter/location combination. However, other constructs which contain different promoters and/or...to study the human GM-CSF promoter and enhancer, a finely regulated promoter controlled by a mixture of...multiple copies of a transcription factor to a promoter and localization of a protein via the presence...are inserted between the truncated gene and the promoter to prevent leaky expression of this resistance...

...cleavage sequence, tRNAs, multiple individual promoters).  spCas9 and its gRNAs are also larger than their...and up to 7 gRNAs Each gRNA requires its own promoter Sakuma et al   Multiplexed Lentiviral Expression..., and up to 4 gRNAs Each gRNA requires its own promoter   Kabadi et al Csy4-Cleavable Cassettes transfection...expressed in 293 cells. This allows for a single promoter to drive expression of a crRNA array and doesn...arrays eliminates the need to include multiple promoters to drive crRNA expression and sequences that allow... Zetsche et al reverses the orientation of the promoter driving expression of the crRNA array (bolded ... Make sure to order 5’ phosphorylated oligos or treat with T4 PNK. Delivery options for Cpf1 multiplexing...

...at cancer cell metabolism. Cancer cells promote oxidative stress leading to the degradation of one key .... Unfortunately, as a whole, microenvironments promote tumor durability instead of repressing them, making...caveolin 1, also known as CAV 1. Usually, caveolin 1 promotes the cell life cycle progression and enhances tumor...signaling, possibly inducing the secretion of growth promoting proteins, such as human growth factor, vascular...Fibroblasts, or CAFs. CAFs are altered fibroblasts and promote the development and metastasis of tumors through...different organs. This means that exosomes not only promote microenvironment’s malignancy but also incite different...tumor’s evasiveness, evolution, and response to treatment. Some microenvironment cells improve tumor’s durability...

...most imporant is that, upstream of the Cbh promoter, this plasmid has a U6 promoter followed by BbsI restriction...SpCas9 This plasmid expresses Streptococcus pyogenes (Sp) Cas9 from a Cbh promoter. Cbh is a modified version...version of CAG promoter that the Zhang lab has tested in a variety of mammalian cell lines. Cas9, of course...

...proteins (bEBPs) at a sequence upstream of the promoter called the upstream activating sequence (UAS). The...σ54-based transcription. Transcription from σ54 promoters requires binding of bacterial enhancer binding...gRNA complex which allows transcription from the promoter.  They found that the system is active in E. coli... and displays high dynamic range. However, the promoter in question must be σ54-dependent and the system...and further the study of natural σ54-dependent promoter systems. Find these CRISPR activation plasmids... reporter by inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter, the researchers...small-molecule probes to aid in the understanding and treatment of cardiovascular disease. Find the LipoGlo plasmids...

...plasmids used to express CRE under neuron-specific promoters (aCaMKII and hSyn). Created with BioRender.com...plasmids used to express Cre under neuron-specific promoters, including calmodulin-dependent kinase II alpha...excitatory neurons) and Synapsin-1 (pan-neuronal) promoters (Figure 1b), and sgRNAs can be found here. Neuron-optimized...expression is mediated by the synapsin 1 (SYN) promoter, which was superior in its ability to induce CRISPRa... CRISPRa in primary rat neurons over other promoters, including ubiquitous ones such as EF-1α and CAG....consequently regulates neuronal physiology by promoting the development of neurons and synapses as well...cells to determine which transcription factors promote neuronal differentiation (Liu et al., 2018). The...

... cauliflower mosaic virus 35S promoter (CaMV 35S) or ubiquitin promoters (Belhaj et al., 2013). Addgene...achieved by placing the gRNA downstream of a plant-specific RNA pol III promoter, such as AtU6, TaU6, OsU6...consistent between plants and other organisms, but the promoter used to drive gRNA expression is dependent upon... gRNA” backbones which contain a plant pol III promoter and gRNA scaffold sequence and allow researchers...is commonly driven by plant-derived RNA pol II promoters which regulate expression of longer RNAs (such...expression). Examples of commonly used RNA pol II promoters for Cas9 expression include the ubiquitously expressing...Summary While different in many of the specifics - promoters used, precise protein sequences or domains, and...

...fairs are also an opportunity for the societies to promote their upcoming events and intro sessions, and give...Since 1996, Cambridge Biosoc’s goal has been to promote interest in the biological sciences amongst the...night. The society also visits high schools to promote chemistry degrees and provide advice to students... to support iGEM alumni-industry networking to promote the creation of start ups and biotech spin offs...Laureates, who speak about their work and current, trending topics in their area of research. The society ...

...tandem with a single promoter, but only the gene directly following the promoter is expressed. Cre-mediated... farther downstream in the construct. As RFP and OFP are directly downstream of the promoter in Brainbow...an RFP, CFP, and YFP downstream of a ubiquitous promoter. Zebrabow is an extremely versatile technology...to neurons by placing Brainbow downstream of a neural-specific promoter, or the entire fish could be labeled...Brainbow system are that it does not use ubiquitous promoters, meaning that not all cell types can be labeled...non-fluorescent YFP and activate fluorescence of the downstream OFP, GFP, or RFP. Cells that express the construct...adult fly. As a stop cassette is located directly upstream of the fluorescent protein genes, Cre recombination...

... the start of the gene to end up just downstream of the promoter found in the backbone. Ideally, the backbone... enzyme cut sites (restriction sites) downstream of the promoter as part of a multiple cloning site (MCS...place your gene in the proper orientation downstream of the promoter, you can add an EcoRI site just 5’ of... leaving the 5’ end of your gene just downstream of the promoter and placing the gene in the proper orientation...easily orient your gene insert with respect to the promoter. For example, let’s say your plasmid backbone ... on the left side of the image below. It has a promoter (blue arrow) followed by the restriction sites...of the backbone plasmid (see below). You should treat your digested backbone plasmid with a phosphatase...

...putting a gene of interest directly downstream of a tissue-specific promoter sounds simpler, this modular system...Gal4 expression can be regulated by any chosen promoter, and Bloomington’s Stock Center has fly lines ...lines expressing Gal4 under a variety of promoters resulting in tissue specific expression. To express YFG... to making a new fly with hedgehog under a new promoter each time a new hypothesis is tested. Controlling...gene expression. Thus, genes and siRNA encoded downstream of the UAS sequence are only expressed when Gal4...systems in Drosophila: A synthesis of time and space, Trends Genet. 20, 384–391 (2004). PubMed PMID: 15262411...I. Bartos, S. Marka, J. C. Kagan, Environmental Stress Causes Lethal Neuro-Trauma during Asymptomatic ...

...bidirectional promoters, or the creation of bicistronic or multicistronic vectors. Unlike promoters which will... proteins from the same mRNA. We've discussed promoters before so in this blog post we’ll cover basics...screenable or selectable markers from a unique promoter, multicistronic plasmids ensure that any cells...originally discovered in poliovirus RNA, where it promotes translation of the viral genome in eukaryotic ... the upstream cistron will have a few additional residues added to the end, while the downstream cistron...scientists have reported lower expression of the downstream cistron due to factors such as the experimental...the end of the 2A sequence and the next peptide downstream.4 The "cleavage" occurs between the Glycine and...

...approximately 19 kb in length. It contains short promoter sequences at its 3’ and 5’ ends that are crucial..., the roles of specific viral sequences (e.g. promoters) can be studied by changing the minigenome cis-acting...minigenome) are under control of a T7 RNA polymerase promoter sequence. Thus, T7 RNA polymerase also needs to...leader region acts as a functional BRSV genome promoter." Journal of virology 73.1 (1999): 251-259. PubMed...Michael, et al. "The Ebola virus genomic replication promoter is bipartite and follows the rule of six." Journal...one developed for Zaire ebola virus (EBOV) are extremely useful, allowing researchers to study aspects ... systems, such as the one described here, are extremely useful research tools. In addition to the reduced...

...vitro models for lung disease. Darrell strives to promote open source biology in his own lab, at the Center...to help keep us focused on our core mission to promote scientific sharing. Listen to learn more about ... defines Open Science and how he practices and promotes open science in the lab - sharing reagents, cell...Darrell's ultimate goal - to develop therapies to treat disease in the lung. Chapter 2: Balancing research...Meidince - Understanding and developing means to treat disease through tissue repair, stem cells, gene ...

...TIR1 of the SCF-TIR1 ubiquitin ligase complex, promoting its interaction with the degron sequence and ultimately...studying the effects of endogenous looping on distal promoters and enhancers, or have attempted to engineer loops...LADL Anchors to genomic targets, such as a distal promoter and an enhancer. What makes LADL unique from previous...group targeted the enhancer region of Klf4 and the promoter region of Zfp462, which are over 500kB apart, ...sequence targeted by the Cas9 nuclease. The Streptococcus pyogenes Cas9 (SpCas9) requires an NGG PAM to...

... gRNA from a Drosophila U6:2 promoter and Cas9 from the actin 5C promoter. Addgene’s fly library web page... over 30 promoters and 140 genes, including constructs with unique tissue-specific promoters expressing...dCAS9-VPR expression is under the control of the TRE promoter, which allows for doxycycline-inducible expression...are cloned downstream of a minimal promoter sequence. Should one of these random stretches of DNA contain...its own transcription from the minimal promoter and its strength can be determined from its enrichment ...many different promoters, RBS’, coding sequences (CDS), and terminators of varying strengths. These can be...fluorophores, generate expression vectors with various promoters suitable for in vivo expression, and/or produce...

...cell-type specific promoters; however, those usually drive weak expression of downstream genes. Strong cell-specific...Optogenetic FLEx vectors contain a strong promoter upstream of an opsin gene (e.g., channelrhodopsin-2...cell-specific promoters, on the other side, are too long for rAAVs, which can only package sequences shorter... the antisense orientation with respect to the promoter to prevent its expression, and is flanked (“floxed...rAAVs expressing Cre under a specific cell-type promoter. Once injected into the brain, the viruses infect...thereby enabling its expression under a strong promoter only in specific neuronal cells. FLEx ensures,...for analyzing the neural circuits of behavior." Trends in cognitive sciences 15.12 (2011): 592-600. PubMed...