...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...

.... Pro-Tip Different brands and lots of FBS can promote or inhibit transfection. Test a variety of brands...control the multiplicity of infection (MOI) in downstream studies. This protocol was developed for A549...Microcentrifuge tubes, Neptune 3745.X 6-well tissue culture treated dish, Corning 3516 15 mL conical tubes, VWR 21008... after 20–30 passages. Do not add penicillin/streptomycin to the media, as it can affect viability and...target cell line needs to be empirically determined. Treat the target cells with a range of doses of antibiotic...vector pRosetta . The following day, cells were treated with puromycin. Colonies were allowed to grow in...EF1α dCas9-KRAB . The following day, cells were treated with puromycin. Colonies were allowed to grow in...

...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...

...However, the same technique can be used to move promoters, selectable markers, or any other DNA element ...single enzyme, but this will require phosphatase treatment of your recipient plasmid as well as a specifically...re-circularization of the recipient plasmid. You should treat your digested recipient plasmid with a phosphatase...

...fluorescent protein eYFP from the hSyn1 promoter Viviana Gradinaru 117383-AAV1 TRE-DIO-eYFP An AAV genome with tet-inducible...-mCherry Expression of hM4D(Gi) using the CD68 promoter. Bryan Roth 75033-AAV9 pAAV CD68-hM4D(Gi)-mCherry...mCherry Expression of hM4D(Gi) using the CD68 promoter. Bryan Roth 83894-AAV1 pAAV-hDlx-Flex-dTomato-Fishell...An AAV genome that expresses tTA from the hSyn1 promoter that contains a positive feedback loop for amplifed...Cre-dependent expression of the tTA from the hSyn1 promoter. Contains a positive feedback loop for amplifed...recombinase-dependent expression of mNeonGreen from the CAG promoter Viviana Gradinaru 100798-AAV1 pAAV-syn-FLEX-splitTVA-EGFP-tTA...expression of the fluorescent protein EYFP from the CAG promoter Viviana Gradinaru 104055-AAV2 pAAV-CAG-eYFP An...

... °C. Pro-Tip Different brands and FBS lots can promote or inhibit transfection. Test a variety of brands...Microcentrifuge tubes, Neptune 3745.X 6-well tissue culture treated dish, Corning 3516 15 mL conical tubes, VWR 21008... after 20-30 passages. Do not add penicillin/streptomycin to the media, as it can affect viability and... reagent for your target cell line. To do this, treat target cells with a range of doses of antibiotic... try higher/lower dilutions depending on your downstream applications. If you’ve titered your virus beforehand...

... 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...

... sensor, he will construct plasmids containing promoter regions that activate fluorescent protein transcription...and to Seeding Labs who helped us coordinate and promote the awards. Additional Resources on the Addgene... has studied various aspects extreme life on earth including extremophile microorganisms, biomarkers, ... to pyrazinamide (PZA) (a drug commonly used to treat tuberculosis) and in identifying new drug targets...biofilm formation. The quest for plant compounds to treat complications of diabetes Gloria Otito Izu, PhD...infusions of L. taraxacifolia leaves are widely used to treat several diseases. Her goal is to identify and isolate...blocked, resulting in reduced blood flow. Novel treatment options, as well as non-pharmacological solutions...

...activated, this DREADD acts through the Gq pathway and promotes neuronal firing. Plasmid Serotype Name Depositor...more accurate cell and tissue targeting. While promoters and AAV serotypes can sometimes be used to target...2-N-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 N protein  pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro...pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro) and NSP2 (pLVX-EF1alpha-SARS-CoV-2-nsp2-2xStrep-IRES-Puro) are also...SARS-CoV-2 M protein pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 ORF3a ...

...expression plasmids in a high copy backbone with a T7 promoter driving multiple combinations of WT, codon optimized...which drive expression using either TEF1 or GAL1 promoters. These vectors are intended for the high level...race to find vaccines, therapeutics, and other treatments to stem the spread of COVID-19. That’s why we...sites (thrombin and TEV), and C-terminal tags (StrepII, 3xFLAG, 6xHIS, MBP, Myc). These constructs include...expression and purification of SARS-CoV-2 proteins for downstream experiments and offer flexibility in tag, cleavage...requested the plasmid. By leveraging our core strengths of intensive screening and the use of high throughput...

...dCas9 along with transcriptional repressors to the promoter region of the gene of interest. dCas9-based repressors... and a dCas9-KRAB fusion, both targeted to the promoter region. Cas9 activators include Cas9 fused to ...the Suntag array of factors, all targeted to the promoter region.   Ok, I have a Cas9! What do i do ... genome.  2) The target is present immediately upstream of a Protospacer Adjacent Motif (PAM), the sequence...

...different promoters followed by leader sequences (encoding small leader peptides) downstream of which ... vectors within this kit contain a variety of promoters to facilitate gene expression in bacteria, yeast...different genes was predictable based solely on the promoters and Shine-Delgarno sequences used in this so-called...transcription from resulting in the production of a downstream peptide, the transcription terminators in this...BIOFAB Plasmid Set contain terminators of varying strengths centered within GFP/RFP bicistronic reporters ...

...targeted gene from its promoter, creating a loss-of-function allele as well as a promoter-FP fusion for the...for the CRISPR/Cas9 revolution!   References Streamlined Genome Engineering with a Self-Excising Drug ...

...unstable fluorescent proteins under the control of promoters responsive to water or nutrient levels can help...fluorescent protein reporter under the control of promoters that control cell cycle gene expression.  Additional...serotonin reuptake inhibitors are frequently used to treat depression and anxiety, but these drugs are often...

...that flank the gene cassette, which contains a promoter, the gene of interest, and a polyA signal. Created...sequences are from AAV2 because they demonstrate high promoter activity (Earley et al. 2020), but non-AAV2 ITRs...limited, but this area of research is important for downstream AAV applications.  Tips & FAQs Due to their high...

...for its powerful promoter of a gene important for methanol usage. This gene promoter, AOX1, was co-opted...Affordable to culture and easy to work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to...

...search scoped to viral preps shows filters for promoter and serotype, among others. We also created a ...example, there is only one viral prep filter for promoter “Syn”, that, when selected, will match variations...members from the Customer Service, Data Curation, Outreach, Product, Scientist, and Software teams. We knew...

...efficient AAV prep for a 4.2 kB insert with the CMV promoter using the AAV-BR1 capsid? A: It is our understanding...4.7kB.  If your insert alone is 4.2 kB and CMV promoter+enhancer is 0.5 kB, then you would be oversized... reliable quantitation. Differences in sample treatment across methods can affect reported titer by either...

...protein whose coding sequence is closest to the promoter will be expressed. This system differs from Brainbow...translation-blocking STOP cassette immediately following the promoter. The STOP cassette includes a mutant YFP that .../FRT recombinase. When placed under different promoters, Brainbow and Flpbow can be used to label distinct...Drosophila. Why Brainbow? Brainbow highlights the trend in biology towards single-cell resolution imaging...

...suitability of various antibiotic resistance markers, promoters, and inducible gene expression systems in bee ...fusion is under control of the arabinose-inducible promoter. Cells are first induced with arabinose to begin...designed to bind and detect nearly any RNA sequence. Downstream of the hairpin is a ribosome binding site (RBS...responds to infection, dysbiosis, and antibiotic treatment. To facilitate these studies, Pamela Silver’s ...

... Atlas (NURSA), one of our primary goals is to promote an understanding of how these processes are regulated...upon a context defined by ligand, cell type, and promoter, organizing the expression of functional groups...transported from cells, traverse the body through the bloodstream, and eventually elicit their actions on other...drugs currently available or being developed to treat a multitude of therapeutic indices, including hypertension...

...either the Thy1 or CAG promoter, while Brainbow AAV is under control of the EF1a promoter. Livet et al., ...known as SpyLigase and is a protein domain that promotes the formation of an isopeptide bond between 2 ...insert fragments of DNA containing basic parts (promoters, UTRs, coding sequences, terminators, etc) into... creating a single transcriptional unit (Ex: a promoter, 5’UTR, coding region, and terminator). Next, ...individual types of pre-cloned insert modules (plant promoter, N-terminal tag, coding sequence of the gene of...Biotech. 2013. Dec 23.   Highly bright and extremely dim GFPs The availability of the three dimensional...unprecedentedly-high brightness, bfloGFPa1, and the other with extremely-dim brightness, bfloGFPc1. The plasmids used for...

...customizable sgRNA from the U6 promoter and puromycin resistance from the EF-1a promoter. It’s a 3rd generation...plasmid from the Zhang lab expresses Cas9 from Streptococcus pyogenes with an invariant sgRNA scaffold and...

...kit contains 96 parts including terminators and promoters, CRISPRi parts, selectable markers, transformation...determined protein expression levels from different promoters, and inhibited gene expression using CRISPRi. ... acceptor vectors from the MoClo Toolkit from Sylvestre Marillonnet’s lab to develop synthetic biology...