We narrowed to 964 results for: Cre

...downstream of the TAM, creating staggered double-strand breaks like those created by Cas12a. Applications...TnpB from Deinococcus radiodurans (Dra2TnpB) to create a genome editing system for plants (Karmakar et... proteins with additional enzymes or domains (increasing the overall size), using TnpB as the core nuclease... using your method of choice. Step 5: Colony Screening Use Primer 92F (5′-cattacgcaattggacgacaac-3′)...′) and Oligo 2 (specific to your target) to screen colonies via PCR. A 354-bp product confirms successful...insertion. Alternatively, use M13R and Oligo 2 to screen. A 667-bp product confirms successful insertion...

...level of antigen in the sample. This figure was created with BioRender.com  Direct ELISAs A direct ... bind non-specifically to assay components and increase background staining. Direct ELISAs also lack the... antibody. Incorporating a secondary antibody increases the sensitivity of the assay, since multiple secondary... both primary and secondary incubation steps, increasing the workload and time commitment for users. This...antibodies for the capture and detection steps, which increases the specificity of the assay. The assay is also...antibody availability Longer protocolLess specificityIncreased chance of cross-reactivity Sandwich ... High specificityHigh sensitivity Longer protocolIncreased chance of cross-reactivityChallenging to develop...

...limitations to facilities and indirect costs, and increased tariffs present an unprecedented challenge to ...to help research institutions maximize their increasingly limited resources: Preserving scientific assets...authenticated materials at a fraction of the cost of creating and verifying them independently — particularly..., and antibody production that labs may find increasingly difficult to maintain or access. These services...with industry and public and private partners to create additional pathways for critical scientific work...community has demonstrated remarkable resilience and creativity in the face of funding challenges. By strengthening...

...was Addgene's first distributed plasmid and was created in the lab of my thesis advisor, Dr. Bruce Spiegelman...requested item of 2005. shRNA - In 2006, we saw an increase in popularity of empty backbones for shRNA expression...requested. Ubiquitin - One particular gene we saw increased interest in from 2008 to 2009 was ubiquitin. The...popular with scientists, but 2012 was marked by increased deposits (and requests) of biosensors, optogenetics...and for so many different purposes that we had to create a CRISPR page to keep up with all the new plasmids...of plasmid sharing? Check out this infographic created by our own Addgenie, Laura Veckerelli. Click on...

...Harvard. There’s an astounding number of ways to create chromosomal mutations in bacteria, so many that...of surrounding genes. However, there are ways to create scarless mutations, ones that don’t leave any undesired..., CRISPR-Cas9 technology has also aided in the creation of scarless mutations, both in stand-alone systems...systems (Jiang et al., 2013), and those which increase the efficiency of Lambda red (Pyne et al., 2015;... recombineering (Khetrapal et al., 2015). They created vectors for Lambda Red that contained one of eight...aeruginosa. These toxins were a mix of Type VI secreted effector toxins, toxin-antitoxin type toxins, ...Enterobacter cloacae, and Serratia marcescens and have created dozens of deletions as well as insertions with ...

...also helps confirm that lanes are loaded evenly. Created with BioRender.com. Loading controls are even...normalized values for the two conditions are equal. Created with BioRender.com. This kind of normalization...must be addressed with separate controls (green). Created with BioRender.com. Researchers often rely ... of the assay is between the two dotted lines. Created with BioRender.com. To see why this S-shaped...protein abundance versus band intensity (right). Created with BioRender.com. On this gel, the protein...loading control that can be used for normalization. Created with BioRender.com. Assumption #1: The loading...protein approach. By eye, it’s difficult to see an increasing concentration of β-actin in panel A. Indeed, ...

...therapeutic interventions for cancer. This screening platform is an incredible boost to the cancer research community...Barcoding (HiTMMoB) technique (Tsang et al., 2016), creating a collection of plasmids each carrying a single...mutations The researchers used the collection to screen individual perturbations in vitro using Ba/F3 and...~22% of the mutations, however, significantly increased cell viability, and these “activators” may be ...annotation of rare gene aberration drivers of pancreatic cancer." Nature communications 7 (2016): 10500...

... in yeast Base editing CRISPRi in Caulobacter crescentus Multiplexed gRNA for human pluripotent stem cells...editing. Using several rapid and cost-effective screening methods, David Liu’s lab identified and validated... base editor plasmids CRISPRi in Caulobacter crescentus Figure 1: Serial dilutions of CRISPRi in...adapted from Guzzo et al., 2020. Caulobacter crescentus is a popular model organism for studying the ...ctrA sgRNA and saw that the levels of ctrA mRNA decreased after 20 minutes in xylose (which controls expression...methylcytosine oxidation to DNA repair, which increases activation of methylation-silenced genes. Read...

.... Figure 1: Diagram of AAV components. Created with BioRender.com. AAV vectors have both distinctive...AAV2, and so on. In addition, researchers have created many engineered serotypes by mixing, matching, ...liver, muscle, lung, CNS, kidney, retinal, and pancreatic cells (Ahuja et al., 2025; Issa et al., 2023;...been shown to transduce skeletal muscle, cardiac, pancreas, smooth muscle, CNS, kidney, and retinal cells...been shown to transduce liver, skeletal muscle, pancreatic, and retinal cells (Issa et al., 2023). AAV11... Kidney AAV8 Liver AAV8, AAV9 Lung AAV6 Pancreas AAV2, AAV8 Photoreceptor cells AAV2, AAV5, AAV8...2025). Evaluation of transduction efficiency in pancreatic beta and alpha cells utilizing various AAV serotypes...

...International Mentorship Program (IMP) USA-Europe was created. IMP as an idea was conceived and developed by ...the specific technical and logistic support to increase the chances of success, but also nurture a supportive...country of origin with an international mentor creates a closer and more approachable relationship for...at IMP In these sessions, mentors helped mentees create “Career Development Plans” which provided paths...developmental needs and resources needed to pursue them, increased their network of contacts important for their ...Conference highlights included remarks by the Spanish Secretary of Research, Development, and Innovation, D. Carmen... as they witness their student’s progress and increases in self confidence. These improvements, combined...

...experiments but we can definitely give you credit for it and increase your impact! Download ReFigure extension...along with the article metadata, legend and user created notes on a “Pinterest for scientists”? ReFigure...Linkedin and ResearchGate profile. To see how to create ReFigures in a few minutes, visit our YouTube [email protected] Please support ReFigure by creating ReFigures and answering this 2 minute survey! ...impact of teaching and doing science is rapid, concrete and open to innovation by all.  Additional Resources...

...positive selection, negative selection, and/or screening. We’re focusing on positive and negative selection...in this blog post, but don’t worry, we’ll cover screens in a future post. Positive selection: Cells that...with or without your inserted DNA) and further screening will help you determine if the plasmid backbone...arises in yeast. For example, Markus Ralser’s lab created a yeast prototrophy kit containing 23 plasmids ...bacteria to grow on sucrose. Plasmid cloning: Creativity with naturally occurring genes  At this point... scientists are finding more and more ways to creatively perform selections in plasmid cloning. Additional...

...are under a Creative Commons 3.0 license, which requires users to give appropriate credit, provide a link...about new scientific techniques and cool plasmids. Creating graphics were usually the last thing I thought..., I found some free design software that let me create graphics that better communicated the science in...reasons why this was the first program I used to create graphics for the Addgene blog. However, it has ...requested, although there’s no guarantee Biorender will create it. Additionally, you can upload your own images...

... biosensor creation, and custom FRET biosensors can be created using the cpFRET kit created by Olivier...Robert Campbell’s lab display calcium-dependent increases in fluorescence of up to 11,000-fold. FRET biosensors...biosensors. Blue flame “cameleon” calcium sensors created by Roger Tsien’s lab contain two fluorescent proteins...calcium binds, it triggers CaM/M13 interaction that increases FRET transfer between the fluorescent proteins...

...membrane depolarization increases the BRET signal whereas hyperpolarization decreases the signal. The authors...technologies that can accelerate your research. The creation of NanoLuc® Luciferase from a rather dull 19kDa... by NanoLuc® luciferase (NLuc) and set about to create an in vivo imaging reporter by linking two CyOFP1...-Lanterns with NLuc and called their improved creations Enhanced Nano-Lanterns. These great imaging tools...the two fluorescent proteins together thereby increasing their FRET signal. FRET is a non-radiative transfer...overlap with this optogenetic tool, they needed to create a calcium sensor with the following characteristics...

...Natsume et al., 2016). Now, the Kanemaki lab has created a new toolkit of plasmids for the N- or C- terminal...makes it easy to tag your protein of interest to create conditional human cell lines. Lastly, the Kanemaki...sites adjacent to an NG PAM in human cells. The increased versatility of the engineered SpCas9-NG and its...enhancers, or have attempted to engineer loops to create genomic interactions that wouldn’t normally occur...develop dCas9 for synthetic looping, the Phillips-Cremins lab developed the light-activated-dynamic-looping...were able to observe loop formation as well as increased Zfp462 expression within 4 hours of blue light...

...self-assembling split GFP (GFP1-10 and GFP11) to increase fluorescence 10-fold after protease activity (... (To et al., 2016). However, the lab wanted to increase the fluorescence range to better study the spatio-temporal...β-strand of GFP, hence the name FlipGFP. FlipGFP increases fluorescence 100-fold compared to other reporters...can be applied to other fluorescent proteins to create an array of multicolor reporters that will enable...apoptosis during embryo development. GFP fluorescence increased over time and showed a clear pattern in the developing... This reporter achieves approximately 100-fold increase in fluorescence after cleavage by executioner ...

...CRISPR pooled gRNA libraries that can be used to screen for genes involved in a wide variety of cellular...processes. While these are widely requested and incredibly useful tools, they require extensive background... infect target cells, and, finally, conduct the screen. To help alleviate some of this labor for our ...optimization-scale infection and at least one screening-scale experiment achieving the gRNA representation...) says: "When you're setting up to do a genetic screen, you need to focus on building a really good model...use along with lentiviral packaging plasmids to create your own lentivirus stocks. Our research scientists...

...David Liu’s lab created the first base editor in 2016 (Komor et al., 2016) and since then has been trying...David Liu’s lab set out to evolve ABEs with an increased compatibility to Cas homologs. Evolving better...bacteriophage, which rely on the expression of gene III to create infectious particles. A plasmid-based genetic circuit...III to base editor activity and editing speed. Created with BioRender.com. Introducing ABE8e - a fast...about 7 times higher than ABE7.10. However, the increased editing speed and targeting scope of ABE8e comes...comes with a price: ABE8e showed increased off-target DNA and off-target RNA activity. To mitigate this...

...could mean needing to increase crosslinking to counteract the effects of increased sample agitation; a ...such as qPCR or next-gen sequencing. ChIP is an incredibly useful application for exploring these interactions...crosslinking, which uses formaldehyde or UV light to create strong but reversible bonds. This will keep the... necessary for most ChIP experiences, but the increased binding can mask the epitopes antibodies recognize...shorter sonication time could be an opportunity to decrease crosslinking. (Homogenization for a tissue sample...