We narrowed to 603 results for: RAN-1
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TypeBlog PostPublishedJune 15, 2017, 2:30 p.m....protein of interest will be problem-free. In figure 1 we provide an example of the OSER assay with a number...can be shared by publishing it as (part of) figure 1, uploading it as a preprint, or documenting it in ...not by Cranfill et al (2016)). Moreover, good performance in the OSER assay is not a guarantee that using...fluorescent proteins when protein tagging is the goal (Cranfill et al, 2016; Constantini et al, 2012). The localization...
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Suggest A Plasmid You'd Like to Find at Addgene
TypeBlog PostPublishedMarch 16, 2016, 5:30 p.m....to reach out to potential depositors in two ways: 1. Fill out our "Suggest a Plasmid" form Fill out the...where we can find the plasmid. While we cannot guarantee that everyone we reach out to will end up depositing... -
Predicting Adverse Reactions to Monoclonal Antibody Drugs
TypeBlog PostPublishedFeb. 13, 2024, 2:15 p.m....immunogenicity score (Mattei et.al., 2022). Figure 1: Generation of immunogenicity scores by predictive...antibodies for the treatment of diseases. J Biomed Sci 27, 1 (2020). https://doi.org/10.1186/s12929-019-0592-z ...ADAs can inactivate the drug and increase drug clearance, lowering its efficacy and increasing the risk... -
Antibodies 101: Stripping and Reprobing Western Blots
TypeBlog PostPublishedAug. 1, 2024, 1:15 p.m....Glycine-HCl 25–200 mM SDS 0.1%–1% (w/v) Tween 20 (optional) 1% (v/v) HCl Adjust to pH 2.0... your gel and membrane transfer as normal. Immediately after transfer, dry the membrane to maximize protein...you from having to run a whole new gel and membrane transfer. Let’s get started! Before you begin In the...manufacturer’s instructions for the membrane you are using. Dry nitrocellulose membranes are brittle, so proceed ... as normal! Stripping your membrane Alright, so you’ve dried your membrane and returned it to buffer, ...’ll want to: Rinse the membrane in water or fresh buffer. Cover the membrane in stripping solution and...a fume hood. Rinse the membrane in water or fresh buffer. Cover the membrane in stripping solution and... -
New Tool for Lineage Tracing: The ClonTracer Library
TypeBlog PostPublishedSept. 22, 2015, 2:30 p.m.... of two outcomes (see schematic depiction below): 1. Resistant cells among the separate replicates would...development of combination therapies. References 1. Hyo-eun, C. Bhang, et al. "Studying clonal dynamics...library was developed when Carrie was a post-doc in Frank Stegmeier’s lab in Novartis Oncology. Bhang did...lentiviral vectors with roughly 73 million semi-random, 30 bp DNA barcodes that are integrated into target... -
CRISPR 101: Cas9 vs. The Other Cas(s)
TypeBlog PostPublishedDec. 8, 2022, 2:15 p.m....none. Best for: targeting single stranded RNA. Table 1: Fast facts on Cas family proteins...The main differences between Cas7-11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity.... it will proceed to generate a blunt DNA double strand break. Cas9 has been studied for genome engineering...an expanding number of others. Best for: a wide range of DNA edits. Cas3 – The shredder What sets Cas3...locates its target sequence and initiates single stranded DNA degradation. Cas3 DNA digestion can proceed...convenient for targeting experiments requiring single strand DNA generation, such as homology directed-repair...therapeutic delivery. Cas14 – Mini, but single strand mighty Unlike many other Cas enzymes, Cas14 targets... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog PostPublishedSept. 16, 2020, 1 p.m....874 . https://doi.org/10.1038/nmeth.3993 Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu.../doi.org/10.1038/s41586-019-0908-x Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani...949. https://doi.org/10.1016/j.cell.2014.02.001 Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz...indel formation and phenotypic changes as early as 1 week post-injection. Livers from these mice were histologically...gene delivery due to their low immunogenicity and range of serotypes allowing preferential infection of ...prime editors. While using AAV to deliver large transgenes might seem daunting, it’s actually a challenge...split AAVs. In general split AAVs break a large transgene in two pieces and package each piece into an individual... -
Hot Plasmids: Bacillus subtilis Libraries, iPSC Reprogramming, CRISPR Tools, & More!
TypeBlog PostPublishedDec. 11, 2017, 2:02 p.m.... Hot plasmids covered in this segment Hot Plasmid 1: B. subtilis knock out and knock down libraries. Listen...on the B. subtilis libraries. Hot Plasmid 2: Transcription factor fusions modulate iPSC reprogramming efficiency... -
Hot Plasmids Podcast Episode 3: Optogenetics Tools, Improved CRISPR Base Editors, & More!
TypeBlog PostPublishedOct. 25, 2017, 1 p.m.... Hot plasmids covered in this segment Hot Plasmid 1: Photocleaveable PhoCl protein for controlling molecular...Hot Plasmid 3: CRISPR base editors with enhanced range and specificity. Listen to to the Segmennt of Improved... -
Hot Plasmids Podcast Episode 2: New RFPs, AAVs, & More
TypeBlog PostPublishedSept. 8, 2017, 7:52 p.m....Hot Plasmid 1: The Tol2 Gateway-Compatible Toolbox for quickly generating zebrafish transgenic lines to ... -
The Future of Research Symposium Boston 2015
TypeBlog PostPublishedOct. 20, 2015, 2:30 p.m....all of its successes, has many deep-rooted flaws (1). As we discovered through the four workshops at the... Follow David on Twitter @driglar. References 1. Alberts, Bruce, et al. "Rescuing US biomedical research...Ambika, et al. "Transforming training to reflect the workforce." Science Translational Medicine (2015)...better solution would be to reduce the number of entrants into PhD programs to more closely reflect the ...symposium. Eve Marder, professor of Biology at Brandeis University, has recently published an article ...Among other involvements, he helped to establish grants through the Sloan foundation for postdoc-related... -
Hot Plasmids and Viral Preps - March 2021
TypeBlog PostPublishedMarch 23, 2021, 1:15 p.m....2021. The AID system relies on a transport inhibitor response 1 (TIR1) expressed under a desired promoter...and accurately in a “single pot.’ Figure 1: Single-Pot Assembly: a) Graphic depiction of chromatin...editing assemblies are sensor-actuators, either transcriptional regulators or chromatin-modifying enzymes. ...cell surface. When the cells interact, GFP is transferred to the receiver cell, and the receiver cell turns...These libraries encode target sites flanked by randomized nucleotides in place of a protospacer adjacent.... Check out the new additions. Image calcium transients in PV interneurons with AAV-S5E2-GCaMP6f, now... -
Participate in the Pets of Addgene Scavenger Hunt
TypeBlog PostPublishedApril 1, 2021, 1:15 p.m....scavenger hunt and the subsequent clue. (Begin with clue 1 below!) You will find five photos this way. At the...Subscribe to the blog to stay in the know! No pranks, just fun and games this April Fool’s Day on the... to find pictures of Addgene pets with Addgene-branded clothing on the blog. Below each photo, the pet... -
How to Prepare for an Industry Interview
TypeBlog PostPublishedJuly 9, 2020, 1:15 p.m....company culture. It’s also a good idea to prepare 1-2 unique questions for each interviewer by looking...to take notes after each interview. Tailor your 1-hour presentation to your audience Start by asking...position, I just had a phone interview. I also transitioned between different lab groups without having ... an opportunity to demonstrate technical and transferable skills that make you qualified for the position...experience less stressful. First, you can make arrangements with those you live with to have a quiet place... -
Plasmids 101: Screening Strategies Used in Plasmid Cloning
TypeBlog PostPublishedSept. 15, 2020, 1:15 p.m....screening blog post for more information. Figure 1: Result of a blue-white screen. Image from Stefan ...Because Sanger sequencing can usually sequence just 1 kb of DNA, custom primers are especially helpful in...information (more on this below). Plating the transformation on ampicillin plates containing X-gal allows... -
The time and cost required to make a plasmid
TypeBlog PostPublishedMarch 23, 2023, 1:15 p.m....plasmid, based on the assumptions outlined in Table 1. Of course, this is for the simple insertion of...any restriction enzymes or vector purchases (Table 1). Restriction enzymes and vector purchases can raise...relevant work in your field! However, there’s no guarantee you’ll get a response on the first try (or any... -
Gendered Innovations: Why Does Sex of the Cell Matter?
TypeBlog PostPublishedJuly 21, 2016, 2:30 p.m....healthcare system was $35.2 billion (or $140 per $1 invested in the trial). The goal of gendered innovations...is the goal of Gendered Innovations. This project: 1) develops state-of-the-art methods of sex and gender...results, but this result shouldn’t be taken for granted for all research going forward. It’s just not that... -
Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens
TypeBlog PostPublishedFeb. 24, 2015, 7:50 p.m....for the transcriptional activation of endogenous genes and it consists of three components: 1. A nucleolytically...complex with guide RNA and target DNA. Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani... sgRNAs for each transcription start site in 15,977 human genes. The transcriptional repressor (CRISPRi...The next insight was to assemble a synthetic transcription activation complex consisting of multiple distinct...together synergistically, modeled after natural transcription activation processes. Konermann and her colleagues...consequences of repressing or inducing individual transcripts. The activator (CRISPRa) sgRNA library uses the...CRISPRi) library contains 10 sgRNAs for each transcription start site in those 15,977 human genes. Weissman... -
Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation
TypeBlog PostPublishedDec. 14, 2021, 2:15 p.m....on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental to daughter strands of DNA....modifications). Ready to get started? Figure 1: Break free from the ball and chain! Epigenetics...Tethering dCas9 to the histone acetyltransferase p300 or the DNA methyltransferase DNMT3A allows researchers...affect transcription independent of their enzymatic activity. They interact with transcriptional machinery...specifically edit methylation sites. Step aside, transcriptionally active dioxygenase When I started to work...by distinct proteins and result in unclear transcriptional consequences (Sprujit 2013). Enzymatic domains...in the genome and directly interacting with transcription machinery. Well, I’ve got some good news... -
CRISPRainbow and Genome Visualization
TypeBlog PostPublishedFeb. 28, 2017, 3:30 p.m....all the colors of the CRISPRainbow. Figure 1: CRISPRainbow was used to track the localization of...is available as a kit from Addgene. References 1. Chen, Baohui, et al. “Dynamic Imaging of Genomic ...spatial relationship of the two genes over time, at a range of distances from 2-75 Mbp. Labeling multiple, intrachromosomal...a system has been previously described for transcriptional activation/repression and genome editing. ...