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...iPSCs. The generation of human iPSCs (hiPSCs) now provides an opportunity to develop and use patient-specific...for reprogramming purposes. In this post, I’ll describe an episomal vector system that increases reprogramming...the CytoTune™-iPS 2.1 Sendai Reprogramming Kit designed to generate clinical grade hiPSCs is sold at $11,000...replication, OriP and the EBNA-1 sequence that encodes for the DNA binding protein EBNA-1 [7]. These two...reported efforts. The pCXLE episomal system thus provides a practical way to generate transgene-free, virus-free...factors they encode Episomal Plasmid Addgene ID# Encodes pCXLE-hSK 27078 SOX2 and KLF4 pCXLE-hUL 27080...

...Cas13a as a diagnostic tool. A pool of DNA or RNA nucleotides containing a sequence of interest (red) is amplified...Reverse-transcription RPA (RT-RPA), respectively. Amplified nucleotides are combined with Cas 13a in complex with crRNA...the target sequence is present in the pool of nucleotides, the nonspecific RNAse activity of Cas13a becomes... sequence is present in the original pool of nucleotides.   What are the potential applications of...can mediate cleavage with spacers from 20-28 nucleotides in length and does not require a specific PFS... the collateral RNA cleavage response is well described in bacterial cells, LwaCas13a does not display...

...site, and titer. Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult...which include calcium and voltage sensors, are the broadest category of viral vector tools, which may help... for vectors in the “controls” category, which includes things like viral vectors that deliver GFP. ...most requested serotypes (Figure 4), our catalog includes vectors in a variety of serotypes, including the...early engagement with UPenn to our launch of the redesigned Data Hub in 2022. It’s influenced the way we ...viral vector educational resources, from eBooks to guides to protocols, which was why I was so excited to...

...features to consider when designing a full gene. A non-exhaustive list includes: Codon abundance relative...that their codons are more appropriate for the desired expression host. This is known as “codon optimization...as easy as it sounds. Even for relatively short peptides, there can be many possible ways to encode them...every amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...PubMed Central PMCID: PMC4794256. This review provides a great overview of codon usage bias 8. Tuller...

...their BioRxiv post, the authors generated and described several improved genetically encoded fluorescent...Takayuki Iwaki, Kimiko Hara, and Kazuo Umemura have described a simple method which alleviates the inclusion...contains the signal peptide of MBP, followed by the desired nanobody (NB) sequence, an AviTAG, and 6xHis tag...expression can vary significantly depending on transgene design and delivery. To overcome these variations in human...Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases... uses CRISPRi to control gene expression. This includes a CRISPRi-based synthetic oscillator, bistable...

...of data. In this two-part blog series, we will describe the importance of bioinformatics at Addgene and...each week. Our internal software was originally designed to help us work in large formats, such as 96-well...Thanks to the flexibility inherent in Nextflow’s design, we were free to use multiple programming languages...alone! The benefits of containers are they are designed to ‘contain,’ in a complete package, all of the... who or which computer ran the process. Another design choice we appreciated about Nextflow was how effortless...small with a print statement and tested that the desired data was available so we felt confident we were...

... that can help with the transition.” The lab-to-desk-job culture shock is a real thing. I experienced...challenging than lab work at times.” Sometimes a desk job is defined as sitting down at a computer and...consider as you make this transition. Of course, a “desk job” could entail any number of career paths--industry...colleague Mike Lemieux told me, “I find that at a desk, it is much easier to stick to timelines. This is...from legal, Diana in finance, and Wu in graphic design. You could dread these interactions, but you could... move away from calling non-academic careers and desk jobs “alternative careers.” Realistically, based...

...scarless mutations, ones that don’t leave any undesired scars in the chromosome. These methods roughly... recombination enzymes can also accomplish the desired genomic modification. This technique is frequently...resolution step (“double crossover”) which leads to the desired mutation and excision of the vector backbone. This...the pDS132 MCS with a versatile, synthetic MCS (designed with the help of the DNA tuner program) (Latynski...purified amplicons upstream and downstream of the desired deletion directly into cut vector with excellent... of interest, from the day primers arrive, the desired chromosomal modification can be made in 5 days....

...site-specific incorporation of phosphoserine (as described below) provides researchers with a valuable tool for understanding...organisms lacked a classic cysteinyl tRNA synthetase, despite abundant incorporation of cysteine in their proteomes...pSer-incorporated proteins on a single plasmid. This includes 4 copies of a tRNA that specifically recognizes...context-specific effects of UAG suppression, as described (6). To determine the stoichiometry of phospho-incorporated...phospho:non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid...

...Wilpitz founder and lead consutant at Experiment Designs Consulting, Inc. “How much do you need [for your...necessary resources. This is why at Experimental Designs Consulting (EDC), we instead have our clients focus...the institution/funding source. Major equipment includes large centrifuges, sequencers and analyzers, and... as many of these tools as possible, this even includes clocks, timers, and phones.  As for reagents and...passion for science. Follow him on Twitter @XpDesignsMGNT        Additional Resources Find other career...the Addgene Blog! Learn more about Experimental Designs Consulting at their website. ...

...R26R-Confetti described in Hugo J. Snippert, et al. (2010) and the MAGIC Marker strategy described in Karine...Brainbow-1 system employs Cre-mediated deletion, the design of Brainbow-2.1 cleverly combines Cre-mediated ...protein sequences are highly homologous, preventing design of antibodies specific to each fluorophore. Third...immediately following the promoter. The STOP cassette includes a mutant YFP that does not fluoresce, but can ...produces a minimum of 8 colors. The use of AAV provides spatial and temporal control without the need ...

...We're excited to roll out the redesigned, revamped, and re-launched Addgene AAV Data Hub, the place [email protected] to learn more. A great submission includes: Images that clearly show viral spread and cellular...your experiment* Functional data, if the report describes a functional vector such as a channelrhodopsin...site as well as the target region, if the report describes a retrograde serotype An abstract that summarizes...work. Sharing your negative data helps others to design better experiments (and wouldn't it be nice if ...

...researchers at Japan’s Hiroshima University who described their new tool in Scientific Reports in June. ... gap. As Sakuma and colleague Takashi Yamamoto describe in their recent study, the construction system...vector-mediated multiplex genome engineering. The system now provides an efficient targeting strategy for multiplex ... validated and ready. “Currently, the kit only includes two versions of Cas9, nuclease and nickase, but...Addgene here. Other Popular CRISPR Posts: How to Design Your gRNA for CRISPR Genome Editing CRISPR-Cas9...

...antibody worked under the conditions described above, and a short description of how the antibody performed....the submitted application, along with a brief description of the experiment. If that data was included ...ELISA — but in all cases, it contains a detailed description of the experimental conditions.     Figure 4...The submission process is easy, with simple, descriptive fields, drop-down boxes when appropriate, and...

... capsid of another. Formally, pseudotypes are described with two numbers separated by a slash: the first...cardiac, and retinal cells (Issa et al., 2023). AAV2 Despite being isolated second, AAV2 is the best studied... choice of AAV vector inherently depends on the design of your experiment. Though we cannot make universal...experimental reports on commonly used vector cargo designs and administration doses. Don't forget to submit...2008). Adeno-Associated Virus (AAV) Serotype 9 Provides Global Cardiac Gene Transfer Superior to AAV1,...., Schaffer, D. V., & Karpova, A. Y. (2016). A Designer AAV Variant Permits Efficient Retrograde Access...

...In 2005, Boyden et al. described the first light-activated tool for controlling neuronal activity, channelrhodopsin... light in combination with retinal) variations designed to give researchers flexibility in using optogenetic...precise timing of the perturbation. Other tools are designed to be activated within a narrower band of wavelengths...function through multiple intra-cellular signaling cascades. Optical methods for controlling GPCR activation...control GPCR activation and downstream signaling cascades (reviewed in Abreu & Levitz, 2020), including ...You’ll need to test for these differences when designing your experiment.  Finding the right information...

...colleagues describe in the PLoS ONE paper that introduced the kit in December, GreenGate includes the six... bring this into your target organism. We have designed a Golden Gate system based on the daily needs ...and cut another,” Lohmann said. “It allows us to design modules that always have the same recognition site...produced will have different overhangs, they can be designed such that they go back together only one way and...

...record breaking month and to find other posts that deserve a second look. 5 Great Apps for Lab Life This very...Stacey from Eastern Tennessee State University and provides a brief rundown of Sean’s top five smartphone ...Jessica Sacher from the University of Alberta and describes a technique you can use to find phage proteins...molecular biology tool, P1 phage. 2) The post describes an ingenious but simple technique to screen for...that, while affecting a small number of people, deserves our attention and has huge potential for the development...

...so he assembled a plasmid kit designed to do exactly that. As described in the journal Science Signaling...When cancers are treated with drugs designed to hit them right where it hurts, the effects are often ...mutant complementary DNAs (cDNAs) representing core nodes that, when over-expressed, constitutively activated...activators for more cancer-related pathways and more nodes within those pathways. With support from the National...

... discusses some of the programs SITN runs and describes some of the more popular topics from the Science... fields as well as students from the ed school, design school, and Kennedy school to discuss the impacts...impacts of science in a social context. Episodes are driven by narrative and story telling. Chapter 5: The...The SITN Blog - Vini and Amy describe how the SITN blog is a source of scientific information for the ...underwater? Chapter 8: Work-Life Balance - Vini describes how she balances working in the lab and working...