...transcription start site, splice sites, exons, and/or promoters to knock out lncRNA function (Hazan and Bester...of introns or the removal of exons. Disrupting promoter or transcription start site function would also...effective in reducing off-target effects since promoter and enhancer sequences remain intact (Engreitz...Local regulation of gene expression by lncRNA promoters, transcription, and splicing. Nature. 2016;539...elements that are crucial for transcription and downstream function. Rather than attempting to cut out the...For example, inserting a polyadenylation signal downstream of the transcription start site will terminate... ncRNAs While CRISPR/Cas9 has proven to be an extremely useful system for modifying and assessing ncRNA...

... elements are cloned into a plasmid upstream of a minimal promoter linked to luciferase. Activation of...enzymatic activity is measured. The effect of promoters or enhancer regions on gene expression can be ...investigate the effect of regulatory elements, such as promoters, enhancers and untranslated regions, or the effect...Cofactors/Substrates Required Not Required Signal Strength Lower Higher Sensitivity Higher Lower Background...regulatory element (wild-type or mutant) is cloned upstream of the luciferase gene in the plasmid, which is...Expression of luciferase can be monitored as a downstream measurement of miRNA efficiency on a target sequence...The gene of interest is cloned into a plasmid downstream of luciferase containing a stop codon, which ...

...realized I wanted a career in science outreach: engaging the public to promote an awareness and understanding...to use to promote future attempts and learned a bit about recruiting scientists to outreach.  Try new ...spray paint to make a trophy for the winner. We promoted the contest for about two months, and we got one...network and graduate school resources daily to promote CINSO events and recruit speakers and participants... executive director of Central Indiana Science Outreach and a Postdoc at Indiana University School of ...founding a nonprofit called Central Indiana Science Outreach, or CINSO. We organize fun science events for ...supportive mentor and graduate office. I cannot stress how important they were to my success as a student...

...Effective science communication is required to promote public support for research and to keep useful ... a graduate student organization that works to promote science communication. Science in the News hosts...and SITN The Most Difficult thing Vini, Amy, and Outreach Scientist Jessica Welch Have Had to Explain Importance...

...plasmids, the MCS is often downstream from a promoter. Insert Gene, promoter or other DNA fragment cloned...cloned into the MCS for further study. Promoter Region Drives transcription of the target gene. Vital...

... a tool that can help you advance your career, promote career growth, and/or switch your field of work...is larger and there are more opportunities for promotion. It’s not uncommon for bachelor’s and master’s..., i.e. Principal Scientist or Scientist I/II. Promotions don’t happen overnight though, and it might take...but in time I realized its value. My master’s strengthened my critical thinking and problem solving abilities...

...design your donor template to include a T7 promoter upstream of the left homology arm, perform in vitro...transcription to create your ssDNA. Note that the T7 promoter will not be included in the final ssDNA template...GG); this design has been shown to increase T7 promoter activity. To avoid re-cutting by Cas9 after editing...containing the homology arms on both sides and the T7 promoter on one end. Typically, Easi-CRISPR works best ...Addgene plasmids. Design PCR primers to add T7 promoter-left homology arm and right homology arm to the...template with 55-100 base homology arms upstream and downstream of the insertion site. If you are using...

... may differ (e.g., a set of various mammalian promoters), the same cloning procedure can be used to incorporate... in this method include fluorescent proteins, promoters, enhancers, polyadenylation signals, inducible...Table 1). Each construct was flanked with a CMV promoter (to drive high-level expression) and a polyA signal...multiple identical polyadenylation signals or promoters Gibson et al., 2009 MXS-Chaining, BioBricks,...and different methods are suited for particular downstream applications. The tradeoffs between various cloning...

...induce transcription at CpG regions of various promoters. Other researchers additionally fused a TAL effector...: dCas9-p300 adds H3K27ac marks to histones in promoter and enhancer regions. These marks are associated...Figure 2: dCas9-Tet1 demethylates cytosines at promoters and enhancers. This targeted demethylation is ...Figure 3: dCas9-DNMT3A methylates cytosines at promoters and enhancers. This targeted methylation is associated...-based acetyltransferase activates genes from promoters and enhancers. Nature Biotechnology 33:510–517...in the DNA oxidation and repair pathways work downstream of Tet1 to restore the DNA after cytosine removal...

...optimism they encapsulate, and the solutions they promote help motivate you to spend a little time advocating...fight is far from over here at home.  The work of promoting and defending the role of science in our democracies... take in abusing climate science skepticism to promote harmful policies that benefit polluting industries.In...marching in and of itself will be enough. IT WON’T! Promoting the incorporation of science in policy, demanding... that uses yeast cells to study how DNA damage promotes cancer. She is also a science communicator, UBC...Hundreds of scientists and supporters in Montreal took to the streets on April 22 to join in the March for...at the Goodman Cancer Research Centre at McGill University in Montreal, Canada.  Outside the lab, she ...

...Regulated Cre expression – placing Cre downstream of promoters that are active only in certain cell or...examined a number of important plasmid elements – promoters, origins of replication, protein tags, and antibiotic...Cre under a variety of ubiquitous and regulated promoters, and many loxP-containing alleles have also been...often called a “lox-stop-lox” or “LSL” cassette) upstream of a gene of interest will prevent gene expression...

...repeat (LTR) of the HIV-1 genome, which acts as a promoter, and found that SAM targeting of the enhancer ...NF-KB binding sites increased activation of HIV promoter-luciferase constructs. In multiple HIV-1 latent...cells with activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can ...indels may not ablate viral function, but rather promote resistance to CRISPR/Cas9. While these results ... acquired immunodeficiency syndrome (AIDS) in untreated individuals. Symptoms of AIDS include rapid weight... viral levels will soon skyrocket back to pre-treatment levels due to the viral reservoirs located in ...in latently infected cells. Although ART-treated patients lack plasma HIV-1, they are at increased risk...

...-inducible promoter, though Khetrapal et al. also demonstrated that the arabinose promoter worked too.... under both rhamnose- and arabinose-inducible promoters. Tight control of toxin expression is absolutely...a rhamnose-based tightly regulated and tunable promoter system." Biotechniques 40.3 (2006): 355-364. PubMed...routinely clone the purified amplicons upstream and downstream of the desired deletion directly into cut...transcriptional terminator BBa_B1002 (from IGEM) upstream of the MCS to reduce transcriptional readthrough...accessibility, selective RNA unfolding and sliding at upstream standby sites." Nucleic acids research42.4 (2013...

...resources and information about organizations promoting reproducibility in this booklet (which was also...line we may see further adoption of policies that promote reproducible publishing practices. One example ... also hopefully see a shift toward hiring and promotion practices that focus on reproducibility and ensuring... Including descriptions of many organizations promoting reproducibility List of Tools for Reproducibility...academia, industry, nonprofits, and publishing. The livestream video from the event can be found below along...interested in, but I’d recommend watching the whole livestream for a more holistic understand of reproducibility...reasons we alter these designs if we do so. The most extreme version of this (and possibly the most effective...

... site accepts the sgRNA target sequence for U6 promoter expression and the second site accepts the homology... a direct fusion of the desired gRNA to the U6 promoter, and a second plasmid containing the homology ...regions of homology to the gene of interest to promote homology directed repair after a CRISPR/Cas9 induced...T2A-TurboGFP-PEST), and small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The Foerstemann... N-terminal tagging vectors (eg GFP, Flag, YFP, Strep, TEV-V5) with either blasticidin or puromycin selection...

...experiment is determined by the capsid and promoter/enhancer strengths in the cell type of interest as well ...transgene expression. Gene regulatory elements [promoters and enhancers, miRNA target sites, and recombination...over systemic delivery. Otherwise, cell-specific promoters and enhancers (Mich et al., 2021) can help restrict...optogenetic and chemogenetic tools. Dual expression Promoters, miRNAs, enhancers, and switches all take up precious...candidate regulatory elements such as enhancers and promoters for cell type-specific targeting (Chan et al.,...when paired with appropriate cell type-specific promoters or Cre lines. Spatial transcriptomics suggests...AAV-PHP.eB, when paired with the ubiquitous CAG promoter, is relatively biased toward L5 and inhibitory...

... repression of transcription through binding a promoter region to inhibit association of transcriptional...targeting their genomic locus of interest, the promoter region of the transcription factor IRF-1. Immunoprecipitation...lab found a microRNA (miR483) that binds to the promoter of the growth factor IGF2 open reading frame to...dCas9, with bound DNA, can then be purified by streptavidin high-affinity purification. This system is both...

...are therefore good temporal reporters to measure promoter activity, for example. Useful positive controls...dark after the transformation. Use introns Plant promoters can be recognized or leaky in Agrobacterium. This...of the GUS gene with the commonly used CaMV 35S promoter in Agrobacterium. This can be easily avoided by...more easily quantify the effect of the different treatments on transformation success. Some reporter gene...

... researchers must be extreme self-starters and especially good at self-promoting—No one is going to tell...diversity and acceptance can be an issue in getting promoted (especially race and gender) Interpersonal issues...especially with peer mentors, to discuss your strengths, weaknesses, interests and passions is another...

...limited by the lack of well characterized inducible promoters. Far removed from the bio-industry’s vision of... the same dCas9 protein. By using an inducible promoter to modulate dCas9 expression, multiple scRNA transcriptional...genes or to introduce heterologous genes, but the trend of research has shifted in recent years toward tinkering...regulators (Zalatan et al. 2015).  These scaffolds are stretches of non-coding RNA interspaced with linkers that...