We narrowed to 792 results for: Por
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TypeBlog PostPublishedJan. 18, 2018, 2:34 p.m.... phage promoter pR, preventing expression of a reporter gene. When light is present, YFI is inactive, ...preventing repressor cI synthesis and allowing reporter gene transcription to take place. Addgene also...promoter is right for me? Here’s a list of some important variables to consider when choosing an inducible...’re expressing a gene that may be toxic, it’s important that your inducible promoter not be too leaky....
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Viral Vectors 101: Viruses as Biological Tools
TypeBlog PostPublishedMarch 21, 2023, 1:15 p.m....permanently integrate into the host genome or temporarily express whatever it encodes. This ability to ...straightforward across applications, while electroporation of a human, for example, isn’t a reasonable...allows you to infect cells and deliver cargo, it’s important to understand their differences to choose the ...PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human immunodeficiency virus type 1 preintegration... -
Q&A with Karen Katz and Carl Paratore, the Newest Members of the Addgene Board of Directors
TypeBlog PostPublishedFeb. 19, 2019, 1:36 p.m....most important resource that Addgene provides the scientific community? Katz: The most important resource...beyond. In your perspective, what do you think are important considerations for the future of open science?...immensely. Paratore: I have two children active in sports, so I enjoy watching their games. If I’m not at... -
Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification
TypeBlog PostPublishedNov. 19, 2019, 2:08 p.m....binding ability of its nanobody partner, so it’s important to insert AsLOV2 at a spot in the nanobody that...light to control signaling provides greater spatiotemporal resolution than pharmacological or genetic ...activated Ras/Erk signaling by using a fluorescent reporter for Erk kinase translocation. These results demonstrate...control protein activity Read about RANbodies, reporter nanobody fusions Learn more about using the secondary... -
Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity
TypeBlog PostPublishedNov. 5, 2019, 1:28 p.m....nuclease for your experiments, there are three important factors to consider: site accessibility, specificity...potential for off-target cleavage should be an important consideration during any experiment. Sensitivity...stringent PAM specificity. This can be especially important for base editing techniques, where Cas9 must be...many options! Comic by Maya Kostman. Another important consideration when choosing a Cas9 is balancing... -
How to Make Friends and Meet People at a Scientific Conference
TypeBlog PostPublishedAug. 7, 2014, 1:58 p.m....scientific conferences. Conferences provide the opportunity to meet people who are interested in the same...the conference. Large conferences have many opportunities but can be overwhelming as far as meeting new...the conference The conference may start in the airport. If you see someone traveling with a poster roll...friendly so stick with those. Don’t miss social opportunities, but don’t overdo it (some of my best studies... -
3 Challenges in Plant Synthetic Biology
TypeBlog PostPublishedJuly 22, 2014, 5:46 p.m....plants are the foundation of life on earth, and opportunities abound to build better fuels, feeds, foods, ...areas in plant biotechnology that are ripe with opportunity. We’ve decided to jump into one of those unexplored...out their mission of creating new forums and opportunities for open innovation in plant synthetic biology...build the plant synthetic biology community and to support more efficient engineering of a better tomorrow... -
Form Your Own Peer Mentoring Group: A How-To Guide for Scientists
TypeBlog PostPublishedFeb. 18, 2014, 4:22 p.m..... But how do you find these elusive teachers, supporters and advisors? I tried to start a more formal ...for their development plans. They all found the support of the group to be intrinsic to their success. ... continue to meet regularly, providing ongoing support, advice and accountability for development goals...mentoring relationship. Celebrate your successes and support each other as you transition to the next stage ... -
Giving gRNAs a Facelift - Synthetic and Beyond
TypeBlog PostPublishedOct. 20, 2022, 1:15 p.m....Nature has evolved ways to prevent degradation of important cellular RNA molecules with post translational...your gRNA. Pro tip: modifications are typically incorporated at the termini of the crRNA molecule (usually...Frontiers in Genome Editing). This is particularly important in experiments in which viability is being compromised...Ramel, MC., et al. Photoswitchable gRNAs for Spatiotemporally Controlled CRSIPR-Cas-based Genomic Regulation... -
Plasmid Tools for Microbiome Studies
TypeBlog PostPublishedJune 27, 2019, 12:25 p.m....microbiome by using engineered genetic tools and reporter microbes. Let’s take a look at a few of these ... because in vitro conditions did not optimally support growth of diverse species. These results further...is a ribosome binding site (RBS) followed by a reporter gene. The hairpin blocks translation by sequestering... reveals the RBS and allows translation of the reporter gene resulting in GFP signal. With the addition... -
Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog PostPublishedMarch 19, 2019, 1:31 p.m....2017). However, Acr inhibition is spatially and temporally ubiquitous, making them a rather blunt instrument...versatility opens up the potential for spatial and temporal control in many genetic targeting and editing ...rapidly screen thousands of proteins for one that can report electrical activity via imaging. They took the ...response to light, offers a way to spatially and temporally control cellular processes in vivo. Many photoactive... -
Rosella: A Fluorescent pH-Biosensor for Studying Autophagy
TypeBlog PostPublishedApril 13, 2017, 2:30 p.m....spectra. DsRed is the permanent fluorescent tag portion of Rosella: it will emit red fluorescence regardless...mechanisms behind autophagy by tracking what’s being transported to the yeast vacuole (i.e. cytosol, mitochondria...2008). Rosella: A fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in...using a single fluorescent biosensor. Scientific Reports,5(1). doi:10.1038/srep12397. PubMed PMID: 26215030... -
Human Germline Editing Using CRISPR
TypeBlog PostPublishedAug. 10, 2017, 2:19 p.m....containing Cas9 and two different gRNAs, and electroporated them with an ssODN repair template in induced...in cases of disease prevention. However, it is important to consider that already-tested CRISPR alternatives...clinical applications could be considered. It is important to consider the question of broader applicability... large step forward in the CRISPR field, it’s important to remember that the study hasn’t achieved any... -
Seven Tips for Using LinkedIn as a Scientist
TypeBlog PostPublishedOct. 1, 2019, 1:21 p.m....people can’t find you on LinkedIn, you will lose opportunities and hiring managers will think it is odd. For... you don’t add a personal note, you waste an opportunity to start things out right. Also, many people ...requests without a personal message and they may report you as spam or “I Don’t Know This Person.” If that...had a chance to proof read. This might not be important for a good friend, but if you are making a professional... -
Hot Plasmids and Viral Preps - March 2021
TypeBlog PostPublishedMarch 23, 2021, 1:15 p.m....length can and should be varied as it can have important implications for the fusion protein effectivity...upon these original tools that allows for both spatiotemporal protein degradation and quantification of this...et al., 2021. The AID system relies on a transport inhibitor response 1 (TIR1) expressed under a ...a desired promoter with a specific spatiotemporal expression pattern and an auxin inducible degron (AID... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog PostPublishedJan. 5, 2021, 2:15 p.m....in a protein. Cells expressing an RFP reporter plasmid for recombinase activity were exposed ...interest. However, FACS is unable to spatially and temporally target single cells. Now, Francois St-Pierre’...way to isolate individual cells with unique spatiotemporal phenotypes from large heterogeneous cultures...introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim’s lab engineered... -
Overcoming the Challenges of Lentiviral Production
TypeBlog PostPublishedAug. 7, 2018, 12:31 p.m....viral Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...third generation packaging systems, this system incorporates the Tet-off transcriptional activation system...number of harvests will all impact the titer. Importantly, optimized conditions for one transfer plasmid...producing lentivirus, it is time to titer. One important consideration when titering a lentivirus is the... -
Hot Plasmids February 2024
TypeBlog PostPublishedFeb. 20, 2024, 2:45 p.m....difficult to genetically edit due to toxicity from electroporation or poor cellular entry with other transfection...PAGE editing with viral sgRNA targeting mCherry reporter (mChe), with co-incubation with assist peptide.... CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons, with...slightly reduced efficacy compared to AA9-X1.1. Importantly, AAV1-X1 is not neutralized by pre-existing antibodies... -
Open Resources and Plasmid Tools For Studying C. elegans
TypeBlog PostPublishedJuly 18, 2019, 12:55 p.m....publication. 3’UTRome The 3’UTRs of mRNA are important for post-transcriptional regulation in C. elegans...combination of these methods provides unique opportunities to develop computational methods to better understand...system allows scientists to both spatially and temporally knock-down proteins of interest. Plasmids for...intersectional strategy using a split intein for refined spatiotemporal transgene control in Caenorhabditis elegans... -
Design Tips for Prime Editing
TypeBlog PostPublishedJan. 23, 2025, 2:15 p.m....that meant rereading many of the original papers reporting various prime editing tools. These papers are ...editing application. And optimizing its sequence is important, as prime editing efficiency is affected by the...nucleotides. For longer templates, it is even more important to test different lengths and sequences, as unintended... cellular surveillance. To limit unintended incorporation of the pegRNA scaffold sequence into the genome...