...microvesicles (approx. 40 - 150 nm diameter) that are secreted from the cellular membrane into the culture medium...AAVs (Schiller et al., 2018). Exosomal vesicles secreted from producer cells also contain cellular macromolecules...open new ways of improving gene delivery. They increase the efficiency of transduction, broaden the use...

...to keep the audience engaged through a computer screen. For the Reproducibility for Everyone Workshop,...author of the poster,” he says.Viewing a poster on a screen is challenging, requiring you to zoom in to different...canceled may be disappointing, but there are many creative ways to adapt and take advantage of the new conference...

... the stem cell community – a community largely created due to Yamanaka’s seminal work. It didn't take... during his keynote address, appeared almost incredulous that the field has exploded so quickly, citing...explained that it took 50 people nearly 2 years to create the first stem cell line that could be used in ...

..., described below, are typically employed when creating stable cell lines and are not used for general...Stable transfection A stable transfection is used to create a population of cells that have fully and successfully...culture media until 24-48 h post transfection when creating stable cell lines. Looking for Mammalian Expression...

...emitted yellow-green light and exhibited 3-5 fold increased brightness over previous BRET probes, with sufficient...injected intravenously with coelenterazine, with increased sensitivity and faster imaging compared to previous... reporter genes for endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent ...

...production. Finally, we are blessed to have an incredibly supportive Board of Directors at Addgene. Our...leadership of Addgene’s co-founder, Melina Fan, we created a new team to push this project forward. The viral...you to the Addgenies who went the extra mile to create this new service. We knew the scientists were waiting...

...communication skills with training programs, which are increasingly available. But think of this as a jumpstart ...possible outcome of a ComSciCon workshop is the creation of a new science communication initiative that...science communication opportunities in your area, create them.  Seek out collaborations with organizations...

...sequence (pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP ...virus, making it ideal for CRISPR pooled library screening experiments. Find lentiCRISPR v2.(If you’ve peeked...site, a pair of annealed oligos can be cloned to create a chimeric guide RNA. Find pX330-U6-Chimeric_BB-CBh-hSpCas9...

... vector and insert are (separately) digested to create 10–12 base “chewed back” overhangs, they can be...bacteria, which will efficiently repair the plasmid. Created with BioRender.com.   SLIC also opened up a...transformed into bacteria, which repair the plasmid. Created with BioRender.com. How does SLIC compare to...

...use small, incremental changes to understand what works and what doesn't. By being creative, thoughtful...each step as you complete it. Yes, this can be incredibly tedious, but it also works, so do it anyway.  ...

...and their GPCR binding partner, Bryan Roth’s lab created an open source resource called PRESTO-TANGO (parallel...parallel receptorome expression and screening via transcriptional output - TANGO) (Kroeze et al., 2015...are ideal candidates for use in chemogenetics. To create chemogenetics receptors, GPCRs were first mutated...

...under cold temperatures, and frigid conditions decrease the potential for microbial contamination. Additionally...Acclimatisation To resolve this impasse, our team decided to create a hybrid system where we could incorporate cold-tolerance...The event also featured a seminar on “Tools to increase scientific reproducibility” by Benoit Giquel, ...

...well-characterized. During this time, we’ve also created a variety of educational resources in an effort...understanding of Addgene’s customer’s needs. Categories We created a categorization of our information in order to...for promoter and serotype, among others. We also created a normalization mechanism to “collapse” variations...

...mutations through homologous recombination. To further increase the utility of CRISPR/Cas9, it will be necessary...that enhance pol III transcription, potentially increasing the levels of gRNA expression. Since tRNAs are... of multiple gRNAs targeting a single gene can increase the likelihood of gene knockout, or multiple gRNAs...

...funding and time. While institutions that focus on creative problem solving like the Howard Hughes Medical...plant research, more initiatives are needed to increase the body of knowledge. Organizations like Open...research and effectively carry out their mission of creating new forums and opportunities for open innovation...

... the world of science publishing, and an ever increasing number of journals and publishers to choose from...them needing a subscription to the journal. An increasing number of funding bodies insist that the research...is beneficial for both the referees (as they get credit for their time and effort) and the scientific community...

... viewed your profile.”  The latter is much less creepy. If we’ve met before, I’d be much more likely to...borders on my first rule of networking--”If it feels creepy, don’t do it.” It’s just not a useful way to build...rules of good resume writing. Include examples of concrete accomplishments in both the technical and “plus...

...cells in the microenvironment. This results in increased expression of transforming growth factor-β signaling...signaling, possibly inducing the secretion of growth promoting proteins, such as human growth factor, vascular... (Kogure et al., 2011) believes that miRNA may decrease the expression of TAK 1, an enzyme that controls...

...publishing has a number of drawbacks, especially the increasingly growing time from submission to publication....for their efforts, inspire future research, and increase their impact. Open Access and Transparency Another...journal publication or an updated preprint, further increasing their findability. Now, go out there and finish...

...electric! After attending this meeting I had an increased appreciation for the role of ion channels and ... the entire worm during regeneration, the worm creates two heads. Subsequent cutting results in a two-...pHLuorin fluorescent protein. This GEVI features increased brightness and improved plasma membrane specificity...

...figure out how to optimize protocols for in vivo screening with lentiviral pooled shRNA libraries. Later,...reagents.” Here at Addgene, we’re excited to see the creative new ways you put the ClonTracer library to use... Blog Learn about the Sabatini and Wood Drug Screening Kit Find Out How to Best Utilize Our Cancer Resource...

...available on our FRET Resource page and can be used to create fusions with a gene of interest. These plasmids...biosensors too, as another laboratory may have already created the sensor that you need. FRET biosensors designed...multiple FRET biosensors simultaneously would further increase the amount of correlated information available...

...response to blue light. This tool, CRY2clust, was created by adding a 9-residue peptide to the C-terminus...regions. To overcome this issue, Shemesh et. al. created a soma-targeted derivative of the opsin CoChR, ...within the cell. Their Fluorescence fLuctuation INcrease by Contact (FLINC)-based biosensors measure fluctuations...

...activation or repression, transgenic mouse line creation, and in vivo gene editing of mice, with the potential...cationic polymer. Polybrene has long been used to increase retroviral transduction efficiency by neutralizing...Nanoblades CRISPR has also been extensively used to create transgenic mice by microinjecting zygotes with ...

...Figure 1: Common functions of natural plasmids. Created with BioRender.com. Resistance (R) plasmids...resistance genes to help cells survive their own secretions. Pro tip! Historically, colicin-producing plasmids...years, and new classifications are still being created as whole-genome sequencing becomes more relevant...

...  Neurogenetic alchemy: CAP-Mac unleashes the secrets of primate brains with brilliance By Hridaya Patel...authors used a multispecies approach to generate and screen a library of AAV9 variants (Chuapoco et al., 2023... Figure 3: Overview of library generation and screening that led to identification and characterization...

...specially engineered sgRNAs to increase transcription. This is done through creating a dCas9/VP64 fusion protein...CRISPRa systems recruit transcription factors to increase gene expression. CRISPRa methods vary in their...

... 2017). In vitro selection allows scientists to create antibodies to targets that may not work well in... laboratories and companies have already begun creating plasmids encoding affinity reagents. These include...open plasmid sharing, the CRISPR community has created hundreds of useful variants of Cas9. By providing...

...using the plexWell technology to easily and quickly create Illumina sequencing libraries. From start to finish...ask the depositing laboratory to review the discrepancies. If the depositing laboratory confirms that ...-Cas9 knockout and transcriptional activation screening. Nat Protoc 12:828–863 . https://doi.org/10.1038...

...growth or decline of interest in a field over time. Creating an alert for a search term helps automate your...is not a traditional search engine. Instead, you create your own research feed by entering keywords, concepts...new papers that contain these terms. You can also create intersects in your feed which means only papers...

...this through a variety of activities, including creating educational materials and lesson plans that are...sterilization of women in marginalized communities have created a legacy of distrust between these communities ...communities, including those by researchers hoping to increase community participation in new precision medicine...

...tips to improve your grant writing and hopefully increase your success rate. 1. Submit your grant early...are unlikely to get past the initial round of screening. I STRONGLY recommend that you ask a PI or mentor...taxes or charitable donations) so there is an increasing realization that, if we want people to keep contributing...

...Mammalian expression plasmids are primarily used to create mRNA and the commonly used mammalian terminators...below) releases the mRNA from the polymerase and creates a free 3' end which is now available for polyadenylation...Internal Polyadenylation Signal Substantially Increases Expression Levels of Lentivirus-Delivered Transgenes...

...Victoria, CA Over the decades science has become increasingly restricted to academic and industrial labs, ...research. This interest manifested itself in the creation of Hackerspaces and Makerspaces back in 2007, ... make accounts with  Addgene. We now have the incredible CRISPR/cas9 tools at our disposal to to modify...

...parallel because the risk of accidental errors increases. In addition to human error, fragments of DNA ...to the reference sequence of the plasmid used to create the viral prep. In a clean viral prep, like AAV...control process! The images in this blog post were created using Snapgene and Geneious software. References...

...disulfide bridges, glycosylation and acetylation to create functional, stable proteins. Protein expression...scientists have taken advantage of the natural secretion of biochemicals and proteins through plant roots... Tagging recombinant proteins with a naturally secreted plant peptide allows for easier access and purification...

...as 30% of all cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations...he or she can use to study it (as seen in the screenshot below). Currently, the page includes interactive... Tackling Cancers' Drug Resistance with a New Screening Kit Learn How the Clon Tracer Library Can Be Used...

.... At Addgene, we use blastn to identify any discrepancies in Sanger sequences, such as mismatches, deletions...determine the potential effect of any nucleotide discrepancies. The blastp and blastx programs are optimized...plasmids instead! All BLAST images are modified screen shots from the NCBI BLAST website. Additional Resources...

...a double-edged sword. An antibody’s large size creates more distance between the target antigen and the...also allows for better tissue penetration and decreases the distance between a fluorescent tag and the...HCab antibodies from its serum, and meticulously screening, optimizing, and characterizing several anti-mouse...

...expense and requirement for lab equipment. Plus they create plastic waste. Kits can also have the annoying ...and filters can be stacked together in series to increase binding capacity. Figure 1: Purify DNA using...Additionally, while commercial plasmid prep kits come in discreet sizes (mini, midi, maxi, giga) which requires ...