...also be a tool for lineage tracing cells that do not tolerate double-strand DNA breaks by introducing ...native endonuclease activity degrades the displaced strand.   Figure 1: EvolvR consists of an error-prone... of PolI3m. This tag would allow researchers to trace the progeny of a single starting cell. Additionally...19.1 (2018): 143. PubMed PMID: 30253789. PubMed Central PMCID: PMC6154929. Halperin, Shakked O., et al....

...Wen et al., 2018). SRIRACCHA uses the Piggybac transposon to stably integrate a reporter gene that contains..., you’re ready to edit. These cells are then transfected with several different constructs for the reporter... gRNAs by simply changing the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The...plasmid.  With this set up, all successfully transfected cells will express RFP while cells containing... Lastly, once a mutant is identified, the PB transposase can easily and seamlessly remove the integrated...

...556.7699 (2018): 57. PubMed PMID: 29512652. PubMed Central PMCID: PMC5951633. Lee, Jungjoon K., et al. "Directed....1 (2018): 3048. PubMed PMID: 30082838. PubMed Central PMCID: PMC6078992. Nishimasu, Hiroshi, et al. "...2018): 1259-1262. PubMed PMID: 30166441. PubMed Central PMCID: PMC6368452. Strecker, Jonathan, et al. "...10.1 (2019): 212. PubMed PMID: 30670702. PubMed Central PMCID: PMC6342934. Teng, Fei, et al. "Repurposing... 4.1 (2018): 63. PubMed PMID: 30510770. PubMed Central PMCID: PMC6255809.  Additional resources on the...

...selection is a process in which only cells with the trait of interest will survive in the selected growth ...unknown manipulation of the population. This is contrary to reverse screens, where there is removal of ...usually selected. The method you choose to assess traits of interest is highly specific to your scientific...the end of the selection process. Non-functional traits will be automatically eliminated, unlike in screens...Directed evolution of enzymes: Library screening strategies. IUBMB Life, 61: 222-228 (2009). doi: https:/...

...brightness When a dish of mammalian cells is transiently transfected with a plasmid containing a fluorescent...experimental conditions that are related to the imaging strategy and the sample. Practical brightness A protein...developed in our lab uses a reference protein that is translated from the same open-reading frame and separated...ranking of practical brightness can be made. This strategy has been used in yeast by Lee et al (2013) and...

...scientists and the support of their technology transfer offices, Addgene would not be able to have such...to recognize a few scientists who have gone the extra mile for this particular project. John Doench and... and careful Addgenies, but also a lot of good tracking. Our Software and IT Teams made major upgrades...inventory management systems to enable efficient, tracked production and distribution of viral samples. The..., a big thank you to the Addgenies who went the extra mile to create this new service. We knew the scientists...

...modeling work in human pluripotent stem cells: a transcription activator-like effector nuclease (TALEN) kit... designer proteins that will introduce a double-strand break in the genome anywhere you would like. Cells...cells after the plasmids have been introduced by transfection or electroporation. All you do to create a protein...not even five years ago, scientists relied on traditional homologous recombination. That technology had...the target site. Also, any time you create double-strand breaks, there is a possibility of introducing a...

...interaction (PPI) studies in plants that rely on transient transfection systems using Agrobacterium (such as Nicotiana...plasmid (Grefen and Blatt, 2012). Similar to traditional Gateway™ cloning, the GOI first needs to be cloned...al., 2016)). During Agrobacterium-mediated DNA transfer, each GOI is usually expressed from a different...plasmids to study PPI using Förster Resonance Energy Transfer (FRET, Heckler et al., 2015) or ratiometric Bimolecular...

... realized that running interactive workshops to train researchers on tools and best practices could be...were covered in the workshop. These workshops illustrate the open and modular nature of the workshop content... post-workshop surveys, recording results, and tracking material downloads. This was particularly helpful...it was realized that interactive workshops that train and provide researchers with tools and best practices...on the Addgene blog: Check out this guide on Transferable Skills: External Collaborations Learn about ...

... the effort associated with a specific topic; extraneous cognitive load refers to the way information ...every project, time, attention, and/or length restraints mean you will have to pick and choose what concepts...summarize the concepts you’re covering. Don't show frustration if they're incorrect; take it instead as valuable...similar misconceptions, it’s time to rework your strategy. Don’t assume it’s the audience’s fault; instead... the information. Maybe I needed to remove a distraction, such as pausing or turning off the slide deck...

...mutations, we check to see if they affect the translated amino acid. We also confirm that the species ...Adeno-Associated Viral Preparations from Viral DNA Extracts. Pooled libraries are a bit more challenging to... work with the depositing lab to develop a PCR strategy suitable for the library in question. We will ...Adeno-Associated Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678 . https://doi.org...2017) Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. Nat Protoc 12:828–863...

...including excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases...recombinase activity. Opto-Cre-Vvd is precise, demonstrating low sensitivity to ambient light, and versatile...-TBN-3’ (where B is G, T, or C). The authors demonstrate that CRISPR-Casɸ is active in vitro as well as... more compact genome editors due to delivery constraints and scientists seeking a wider range of targetable...-based Dual-fluorescent DSB Repair) is a double-strand break repair assay for mammalian cells. It's based...

...genetics in the media (both traditional and social); and synthesizing and translating all these into social ...number of different opportunities to broaden my training beyond the biological sciences, as well as to ...communication experience. These included taking extra coursework and attending seminars in bioethics, ...Find all of our science communication posts Extracurricular activities for a strong science career path...

...fiber optic for the implant depends on the dorsal/ventral location of your region of stimulation (the z-axis...photons to become activated. Light is able to penetrate brain tissue a short distance so ideal implant...jbc.M111.253674. PubMed PMID: 21628464; PubMed Central PMCID: PMC3138296. 2. Sparta DR, et al. Construction...0471142301.ns0216s58. PubMed PMID: 23042500; PubMed Central PMCID: PMC3466483. 4. Kim TI, et al. Injectable...science.1232437. PubMed PMID: 23580530; PubMed Central PMCID: PMC3769938. 5. Iwai Y, et al. A simple head-mountable...

...your insert, DpnI digest away the templates, and transform the PCR products into E. coli. Ligases within ...Otherwise you let your E. coli do the work for you. Traditional restriction enzyme and ligase based cloning is...at a price of a few dollars per reaction for traditional restriction cloning. Sequence and Ligation Independent...to pick the right enzymes, optimize reagent concentration, or purify DNA fragments from gels. If you’re...11.1 (2011): 92. PubMed PMID: 21992524. PubMed Central PMCID: PMC3207894.  Additional Resources on the...

...generate a classic double-stranded break, while modulators regulate transcription and chimeric efforts make... community. The rapid adoption of new tools, illustrated in Figure 2, and the longevity of tools seen ...FR=France; KR=South Korea; CH=Switzerland; AU=Australia.  It probably comes as no surprise that the...plasmids. But many other countries, from Japan to Australia, are significant depositors and requestors. CRISPR...

... lab has created a novel nanocompartment for transporting proteins in yeast by using murine polyomavirus...providing enzyme protection while still allowing substrates and products to diffuse freely into or out of...photocleavable gRNAs within seconds. A CRISPR-based lineage tracing tool captures rare clones that represent 0.001%...where gRNA expression is induced by anhydrous tetracycline. Three new anti-CRISPRs were identified that...expressing GCaMP6f with this promoter. Image calcium transients in PV interneurons with AAV-S5E2-GCaMP6f are ...

...developing generic strategies for detecting cloned genes (Munro and Pelham, 1984). The strategy they landed ... available - aka peptide tagging.  Key to this strategy was the existence of high quality antibodies for...an amide group added to its C-terminus post-translationally, which turned out to be critical for antibody...that Munro and Pelham were devising their tagging strategy, another group of researchers was working on isolating...

...financial and logistical stress that accompanies traveling for campus visits. This is definitely an advantage...lab, research, and provide them with a CV or transcripts if requested Thank them, and eagerly await a ...financial aspects that are confusing? During a traditional campus visit, many students will have questions...the online format, I didn’t have to worry about traveling during an already stressful time, and I was more...universities change their format of interview from a travel day to an online call in response to challenges...

... work has always been focused on gene transfer, from transgenes to plants, chloroplast to the nucleus,...well in plants and give similar stability to transcripts as the ones that people use all the time. Now...coding sequence to the MoClo system. It’s quite straightforward really and we’ll be producing an instructional...Plant SynBio Reading: Quick, Versatile Plant Transgenesis with GreenGate Plasmids  ...