We narrowed to 603 results for: RAN-1
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TypeBlog PostPublishedDec. 30, 2015, 3:30 p.m....forward to another great year of sharing! References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout... name suggests, lentiCRISPR v2 is a lentiviral transfer vector used to deliver both Cas9 and your gRNA...metalloproteinase-9 secretion, and identifying lysine methyltransferase substrates. We’re incredibly happy that lentiCRISPR...
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Why Do I Need an MTA Anyway?
TypeBlog PostPublishedMay 1, 2014, 8:23 p.m....is here to help. Contact us at [email protected] or +1 617-225-9000, or check out our MTA reference pages...Uniform Biological Material Transfer Agreement (UBMTA) for its material transfers. The UBMTA was drafted by...organizations Assurance that recipient may not use the material for a commercial purpose Assurance that recipient... has time for paperwork? Here’s why a Material Transfer Agreement (MTA) matters: Science thrives on collaboration...universities and institutions require them for material transfers. When Addgene entered the plasmid sharing space...receiving institution. Therefore, many technology transfer officers are familiar with the UBMTA and are happy... material transfer agreement... -
How Exercise Purges Stress and Keeps Depression at Bay
TypeBlog PostPublishedNov. 25, 2014, 4:02 p.m....’ve come to the right place. Get started with PGC-1 plasmids from Addgene. References: Skeletal muscle...PGC-1a1, a transcriptional coactivator induced in skeletal muscle by exercise and endurance, protects ...have reported a mechanism based on studies of transgenic mice that helps to explain how it works. The ... skeletal muscle expression of kynurenine aminotransferases, to enhance the conversion of kynurenine into... -
Making Career Development an Integral Part of the Science Training System
TypeBlog PostPublishedFeb. 25, 2021, 2:15 p.m.... successful after their training. Figure 1: Professional Development Hub Workshop Attendees and...ample resources and a positive perspective on the transition to a post-academic career. This training was ... inadequate job of supporting science trainee transition into diverse, successful careers. We’re not ...components of predoctoral and postdoctoral training. A grant from the National Institute of General Medical Sciences... -
Designer PUF Proteins for Any RNA Target
TypeBlog PostPublishedJune 17, 2014, 7:57 p.m.... specific post-transcriptional regulation of endogenous RNA. J Biol Eng. 2014 Mar 1;8(1):7. doi: 10.1186...ability to direct enzymes – from nucleases to transcription factors – to specific sequences of DNA has become...Biological Engineering in March is fused to a translational repressor. In principle, however, designer PUF... -
Hot Biosensors 2022: Year-End Roundup
TypeBlog PostPublishedDec. 6, 2022, 2:15 p.m....improved sensors for these and other targets. Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted...fluorescent biosensors. Nat Commun. 2022 May 25;13(1):2919. doi: 10.1038/s41467-022-30685-x. Feel the ...new class of sensors for neuropeptides and neurotransmitters. Motivated to develop a biosensor for oxytocin...devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA...sensors based on GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost ...kinetics and localization for imaging synaptic transmission. bioRxiv 2022.02.13.480251. doi: 10.1101/2022.02.13.480251...2022.02.13.480251 JEDI-2P voltage sensor The François St-Pierre lab and colleagues have developed a new... -
CRISPR-Cas14: a family of small DNA-targeting enzymes enabling high-fidelity SNP genotyping
TypeBlog PostPublishedNov. 29, 2018, 1:53 p.m....infections, cancer, and other diseases. References 1. Harrington, Lucas B., et al. "Programmed DNA destruction...that Cas14 could indiscriminately cleave ssDNA in trans like Cas13 and Cas12 do for RNA and dsDNA, respectively...can cleave fluorescent probes of dsDNA and RNA in trans. Fluorescent signal indicates the presence of this... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog PostPublishedAug. 23, 2018, 12:05 p.m....fluorescent in acidic organelles (Figure 1), showing that its acid tolerance is maintained in cells. GFP is ...References 1. Zacharias, David A., et al. "Partitioning of lipid-modified monomeric GFPs into membrane microdomains...of different fluorescent protein are available (Cranfill et al., 2016 and FPbase.org). A number of variants...specific RNA stem loop-structure (Querido and Chartrand, 2008). Finally, a nuclease-deficient Cas9 can...clearly the case when proteins undergo post-translational modification with fatty acyl chains. For instance...28360214. PubMed Central PMCID: PMC5385933. 6. Cranfill, Paula J., et al. "Quantitative assessment of ...PMC2644460. 12. Querido, Emmanuelle, and Pascal Chartrand. "Using fluorescent proteins to study mRNA trafficking... -
March for Science
TypeBlog PostPublishedApril 21, 2017, 2:30 p.m....education, government, and civilization in general (1). Science and politics? Science is an apolitical process...consequences of a march. Three of these risks include: 1) Lost trust Perhaps the greatest risk is that the ...working full-time, and have received a research grant in the last five years as polled in the American...climate change page on whitehouse.gov, freezing grants, and proposing to defund aspects of research at...following subjects: Climate change Vaccines GMOs Transgender issues Reproductive health Immigration Pollution...while many also seek to honor and credit women, immigrants, and minorities who have had great impacts on...encouraged by “Broader Impacts” requirements for certain grants) (28). As one young scientist, Matthew Niederhuber... -
CRISPR 101: Validating Your Genome Edit
TypeBlog PostPublishedNov. 3, 2022, 12:15 p.m....by insertion and deletion frequency. Fig. 1: Overview of TIDE. The input is sequencing trace files...keep your product to knock-in size ratio below 10:1. Small knock-ins Is your knock-in edit small, with...optimizing this step, or consider enriching for transfected cells by flow sorting or drug selection, assuming... -
Antibodies 101: Introduction to Gating in Flow Cytometry
TypeBlog PostPublishedApril 4, 2024, 1:15 p.m....at a low angle (FSC) or large angle (SSC) (Figure 1). FSC values depend on the cell's size, while SSC ... or ask experienced colleagues. Figure 1: The laser pulse illuminates the cell and is scattered...software in order to generate signals in a reasonable range. If colleagues in your group already have experience...experimentally-specific gates. Two-dimensional staining (quadrant gating) For our last example plot, I will show...cells (CD8+) from dendritic cells (CD11b+) using a quadrant gate. Within my “lymphocyte + single cell + live... two stains in relation to each other. Using a quadrant gate, I can divide my cell population into DN,...for each stain respectively). (B) Instead of a quadrant gate, I am free to use multiple individual gates... -
Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods
TypeBlog PostPublishedMarch 1, 2016, 3:30 p.m....with the Gibson assembly master mix for 1 hour, and then transform into competent cells. The success rate...you should consider giving it a try! References: 1. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison...generating long overhangs which allows the single stranded regions with homology to anneal a polymerase, ...hour or so, the sample is immediately ready to transform into competent cells. The master mix of enzymes...if the ends of the fragments have stable single stranded DNA secondary structure, such as a hairpin or ...would directly compete with the required single-stranded annealing and priming of neighboring assembly ... -
A Control for All Seasons
TypeBlog PostPublishedOct. 5, 2023, 1:15 p.m....compare the experimental group. Figure 1: A, Most biological experiments will benefit from ... Histochemistry & Cytochemistry, vol. 59, no. 1, SAGE Publications, Jan. 2011, pp. 6–12. Crossref,...across samples and equal protein transfer from the gel to the membrane. A loading control is critical for...identically, used to assess naturally occurring random biological variability. If one were studying the...purple) and nonspecifically through the Fc region (orange). The Preimmune serum control will not bind to .... Always probe the loading control on the same membrane that the target protein was measured on. Flow... -
CRISPR 101: Off-Target Effects
TypeBlog PostPublishedFeb. 8, 2024, 2:15 p.m....downstream experiments. You only successfully generate 1 null clone and proceed with it. In this case, the ...there are 10 putative off-target locations you can 1) determine if editing at those sites would confound...While this may effectively serve as a transfection or transduction control, a different gRNA would have..., you’ll want to select a Cas enzyme with low tolerance for mismatches within the gRNA sequence. ...near each other, this will result in a DNA double strand break (DSB), the event required for frameshift ... -
Design Tips for Prime Editing
TypeBlog PostPublishedJan. 23, 2025, 2:15 p.m.... and a reverse transcriptase (RT) template including the desired edit. Figure 1: Prime Editing ...doi.org/10.1038/s41587-021-01039-7 Nishimasu, H., Ran, F. A., Hsu, P. D., Konermann, S., Shehata, S. I....nicking of the unedited strand and encourage the cell to use the edited strand as a template during DNA...on a fusion enzyme of Cas9 nickase + reverse transcriptase and a prime editing guide RNA (pegRNA). The ...although sequences with G/C content outside this range can still be optimized (Anzalone et al., 2019). .... Test different lengths of reverse transcriptase template, starting with about 10–16 nucleotides. For...pegRNA primer binding site (PBS) and reverse transcriptase template (RTT). Created with BioRender.com. ... -
Bioinformatics at Addgene
TypeBlog PostPublishedJuly 14, 2022, 1:15 p.m.... same output regardless of who or which computer ran the process. Another design choice we appreciated...drives our bioinformatics needs Fig. 1: QC'ing data at Addgene is like standing under a Niagara...automating the bioinformatics steps needed to transform the raw sequencing data into a complete plasmid...appropriate place in our workflow without having to translate or convert programs to a uniform language. The...concepts and vocabulary of Nextflow so that we could translate our pipeline steps into processes and then connect... -
Fluorescent Proteins 101: History of Fluorescent Proteins
TypeBlog PostPublishedAug. 7, 2017, 1:58 p.m....in the crystal jellyfish Aequorea victoria (Figure 1) in molecular detail. On the underside of the “umbrella...glowing protein could be used to measure gene transcription, as well as to track protein localization. And...bioluminescent, some types of corals and anemones are vibrantly colored. He hypothesized that GFP-like proteins... -
Top 10 Open Science Developments of 2013
TypeBlog PostPublishedDec. 19, 2013, 3:46 p.m.... science 1) In February, the Office of Science and Technology Policy issued a memorandum calling ...(66 percent) gave preference to journal access granted via a public repository or authors’ own web sites... -
Our Most Popular Blog Posts from 2019
TypeBlog PostPublishedDec. 31, 2019, 2:26 p.m....changes. 1. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection by Alyssa...editing. Using a fusion between Cas9 and reverse transcriptase and a prime editing guide RNA, it can introduce... -
Bringing Sustainable Practices to the Lab: Innovations
TypeBlog PostPublishedMarch 7, 2019, 2:39 p.m....Consistency, and Transparency. This label scores products by their environmental impact on a scale of 1-10: the... but with the right incentives (for example, by grant agencies), we can shift toward a more sustainable...your RMV to the landfill, it’s collected and transferred to the Red2Green processing facility, where it...