We narrowed to 1,141 results for: Gene
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TypeBlog Post...from Jiazhang Lian’s lab that enables simultaneous gene activation, repression, and editing with a single...studies. Duong et al., Genetics 2021. https://doi.org/10.1093/genetics/iyab006 The CRISPR ... need. Here's what you'll find in this post: Epigenetic engineering with single-pot assembly Tracking...CRISPR plasmids New items from the viral service Epigenetic engineering with single-pot assembly By Eric ... of plasmids to the repository that will help epigenetic researchers custom design their own engineering...) Golden Gate assembly (backbones available at Addgene) with examples of potential synthetic chromatin...with BFP, sender cells expressing surface BFP generated BFP-labeled receiver cells. What’s more, G-baToN...
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Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9
TypeBlog Post...have been modified for genome editing, activating gene expression, visualizing genomic loci, and much more...plasmids from this publication at Addgene. Resources on the Addgene Blog Read about Cas9 Homolog Cpf1...effect on specificity was similar; SpCas9-HF1 generated fewer off-target cuts when compared to WT SpCas9...collaboration with the Joung Lab, they used targeted mutagenesis to create HypaCas9 (SpCas9 N692A/M694A/Q695A/...-HF1. The HypaCas9 plasmids are available from Addgene here! Future possibilities for Cas proteins With...26628643. Find plasmids from this publication at Addgene. 2. Nishimasu, Hiroshi, et al. "Crystal structure...26735016. Find plasmids from this publication at Addgene. 5. Chen, JS, et al. "Enhanced proofreading governs... -
Rinehart Lab Phosphoprotein Reagents
TypeCollection...useful for the incorporation phosphoserine into your gene of interest....by adding this phosphorylated amino acid to the genetic code of E. coli (Park et al., Science 2011). Follow... in Mohler et al., Mol Syst Biol 2023 and find general tips on phosphoprotein expression in rEcoli XpS... Biotech. 2018). This phosphosite library was generated in a single mixed pool by expressing and purifying...phosphosite library ( 111704 ) is available from Addgene. This library is anticipated to be useful for laboratories...assessments. The mode #1 phosphosite library can be generated with either phosphoserine using SepOTSλ ( 68292...domain of NEDD4-2; 111705-8 ) is available from Addgene. Screening kinases. The mode #1 phosphosite library... -
Plasmids 101: Protein Expression
TypeBlog Post...cells) that allow for continuous expression of genes integrated into the insect cell genome. Both of ...and cell free expression systems. Overall the general strategy for protein expression consists of transfecting...systems Yeast are a great expression system to generate large quantities of recombinant eukaryotic proteins... species due to its use as a model organism in genetics and biochemistry. When using S. cerevisiae, researchers...large-scale, stable production of proteins. In general yeast expression systems are easier and cheaper... use. Alyssa D. Cecchetelli is a Scientist at Addgene. She received her PhD from Northeastern University...proteins in plant root exudates Resources on the Addgene Blog Learn about inducible promoters Learn about... -
Plasmids 101: Optimizing Plasmid Yields
TypeBlog Post...fewer transcripts of their antibiotic resistance genes and can therefore be cultured in media containing...plasmid prepping is a breeze. You get your stab from Addgene, streak for single colonies, sub-culture, and prep...use a different strain. For most applications, Addgene uses and recommends DH5𝝰! This strain consistently...density and plasmid yield as these factors are generally, but not perfectly, correlated. Typical growth...sufficient gas exchange in the overnight culture. Generally, large cultures in flasks may be shaken around... strain instructions from the manufacturer. On Addgene plasmid pages, you may see depositor recommendations...writing of this post. Additional Resources on the Addgene Blog Visit our Plasmids 101 Topic Page Learn How... -
CRISPR Plasmids - Empty gRNA Vectors
TypeCollection...are expressed as polycistronic glycine tRNA-gRNA genes (PTGs) and then PTG cassettes are assembled into...capable of simultaneously repressing multiple target genes in Escherichia coli. Koffas Do you have suggestions...BsaI none S. pyogenes Chloramphenicol Marraffini pCas9 42876 Bacteria BsaI yes, cut S. pyogenes Chloramphenicol...none S. pyogenes Qi pDD162 (Peft-3::Cas9 + Empty sgRNA) 47549 C. elegans yes, cut S. pyogenes Goldstein...elegans BsaI none S. pyogenes Joung pCFD3-dU6:3gRNA 49410 Drosophila BbsI none S. pyogenes Virmilion Bullock..., cut S. pyogenes Puro Liu pCFD4-U6:1_U6:3tandemgRNAs 49411 Drosophila BbsI none S. pyogenes Virmilion...Mammalian AfIII none S. pyogenes Bleocin Church MLM3636 43860 Mammalian BsmBI none S. pyogenes Joung pSPgRNA 47108... -
Technical Design of a Western Blot
TypeBlog Post...error than normalizing to so-called housekeeping genes (Aldridge et al., 2008). Woah! That was a lot of...quantification Normalizing to ubiquitously expressed genes Not as accurate; increased antibody costs; increased...Antibodies Resources on Addgene.org Addgene's Western Blot Protocol Addgene's Western Blot Protocol Video...Looking for a western blot protocol? Check out Addgene's western blot protocol and protocol video! Antibodies...gel. Handcast gels are significantly cheaper and generate less waste. However, they also add time to an ...in size. However, these systems are expensive, generate plastic waste, and limit the amount of optimization... can only compare samples to a standard curve generated on the same blot. If your protein of interest ... -
Antibody Guide
TypeCollection...Recombinant antibodies are created by cloning antibody genes from specific hybridoma cells into plasmids. Plasmids... tubulin (sometimes referred to as housekeeping genes). This allows for relative comparison of expression...Science Guides Antibody Guide Antibody Guide View Addgene's Antibody Collection Jump To: Antibody Guide Production...with myeloma cells (immortalized B cells). The genetic reorganization that creates variety in the antibody... be produced from plasmids, with an antibody’s genetic code inserted into the plasmid. This produces the...antibody is the antigen binding site. In B cells, the genetic region responsible for the variable region goes...epitopes on one antigen. Monoclonal antibodies are generated from cultured B cells, where each culture should... -
How to Write a Scientific Review Article
TypeBlog Post...T-cell development, physiology and malignancy." Genes & cancer 6.5-6 (2015): 184. PubMed PMID: 26124919...PMCID: PMC4482241. Additional Resources on the Addgene Blog Writing Scientific Manuscripts: Literature...Manuscript 6 Tips for Grant Writing Resources on Addgene.org Check out Our Hot Plasmids Articles to See Compiled... -
Mesothelioma - Causes, Symptoms, and Treatment
TypeBlog Post...studies show that certain mutations of the BAP1 gene play a role in susceptibility to developing mesothelioma...Additional Resources on the Addgene Blog Find out More About Cancer and Addgene's Cancer Collection Pages...others never do, but we do know that there are some genetic factors involved. For example, multiple studies...research is being done to see what, if any, other genetic factors may be in play. However, there is no question...options: Early detection, immunotherapy, and angiogenesis inhibitors New research is also going into two...bevacizumab (Avastin). This drug (known as an angiogenesis inhibitor) prevents the growth of new blood ...the use of asbestos, thus ensuring that future generations will be protected from this dangerous material... -
SciComm: Taking Science to Elementary and Middle Schools
TypeBlog Post... terms of the vocabulary--DNA, protein, plasmid, gene, etc. Though I talked about CRISPR’s molecular mechanism...quick tips to get things going: If your PTA has a general meeting and a means to request funding for events...7th grade biology teacher. I led up to my job at Addgene, which is likely a career path none of them had...offered engaging examples of how understanding genetics is so timely.” Genome engineering, and many other...guidance and feedback. Additional resources on the Addgene blog Browse our CRISPR blog posts Download our...science communication blog posts Resources on Addgene.org Check out our comprehensive CRISPR guide Find... -
A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog Post...Calabrese (activation), from Addgene to better understand which human genes are important for SARS-CoV-...deposited at Addgene. Plasmids deposited from Alejandro Balazs’s lab includes those for generating pseudovirus... deposited the plasmids used for this study at Addgene which include a C-terminal deletion in the spike... Bst polymerase constructs from their assay to Addgene. Drew Endy’s and Philippa Marrack’s labs also ...investigate how the virus interacts with human cells. Addgene’s mammalian target plasmids express these and other...the COVID-19 pandemic, the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open ...10.1089/crispr.2020.0030 Additional resources on the Addgene blog Learn more about mapping the SARS-CoV-2 -... -
Hot Plasmids - November 2023
TypeBlog Post...delivery of genetic cargo By Alyssa NeuhausTo improve delivery and reduce off-target effects of gene therapy...system to improve on-target delivery of genetic cargo Optogenetic proximity labeling and TransitID Tracking...continuous evolution and protein engineering to generate seven new prime editors, PE6a–g, that overcome...10.1038/s41467-023-40788-8. PMID 37612276. Optogenetic proximity labeling and TransitID By Mike Lacy...transcriptional history of a cell. Lin et al. generated a fusion protein called iPAK4 that assembles into...43R] has been one of the hottest antibodies in Addgene’s collection since we began distribution and has...[K28/43R] under the Application section of the Addgene antibody page, and learn more about the whole-brain... -
PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs
TypeBlog Post...to make sense of biology, with a special focus on gene editing techniques. References and resources References... versatile genome manipulation. Nature Reviews Genetics, 24(3), Article 3. https://doi.org/10.1038/s41576...DeepSpCas9, a deep learning–based model with high generalization performance. Science Advances, 5(11), eaax9249...). Chromatin context-dependent regulation and epigenetic manipulation of prime editing. Cell. https://...j.cell.2023.03.034 Resources More resources on the Addgene blog How to Design Your gRNA for CRISPR Genome ...and Flexibility to CRISPR Editing Resources on Addgene.org CRISPR: PRIME Edit Collection The CRISPR Guide... -
Course-Based Undergraduate Research Experiences (CUREs)
TypeBlog Post...my students published their work identifying new genes required for stem and progenitor cell maintenance...incorporated a discovery-based lab on planarian regeneration into one of my own courses, I was excited to...be authentic – students should be attempting to generate new knowledge that will be of genuine interest...Provost’s course recently used a combination of phylogenetic analysis, structural prediction, and bench work...recombinant enzymes in bacteria, purified them to near homogeneity using a 6XHis tag, and compared their specific... -
Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity
TypeBlog Post...the expected complete sequence for the lacI/lacZ genes (1), while sequencing of white colonies showed a...PMCID: PMC6061786. Additional resources on the Addgene blog Learn the basics of Golden Gate assembly ...other plasmid cloning techniques Resources on Addgene.org Find basic molecular biology protocols Find ... -
Career Insights: Technical Support Specialist
TypeBlog Post...in the fields of gene and cellular therapies. Additional Resources on the Addgene Blog Read What ...Technical Support Calls Resources at Addgene Visit the Addgene Careers Page Check Out Our Career Videos...You will also help troubleshoot products, give general experimental guidance, and resolve complaints about...willingness to help others with their problems, a general positive attitude, and an open-minded personality...will do well in a technical support role. What general characteristics do jobs in technical support have...covered by the lab based team as part of their general job duties. Bigger companies have dedicated technical...post. Klaus Wanisch is an Outreach Scientist at Addgene. These days he's doing a lot of science communication... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog Post...from transient overexpression of protein fusions to gene-edited cells can decrease the number of fluorophores...Fluorescent Protein Should I Use? Resources on Addgene.org Generate Fluorescent Protein Fusions Request Plasmids...in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion with your... dye ligand (e.g., FlAsH, ReAsH) and a short genetically encoded tetracysteine (Cys4) peptide tag could...TMP-tag) – These widely used systems consist of a genetically encoded enzyme variant tag that reacts specifically...such as paclitaxel. Unlike other systems, no genetically encoded tag is required as the small-molecule...strategies have trade-offs between the size of the genetically encoded tag, the speed and selectivity of the... -
Tips from the Repository Trenches: Using Barcodes to Track Samples
TypeBlog Post...contain either an ampicillin or kanamycin resistance gene. Every step along the way involves good aseptic ...of tube for each plasmid we have at Addgene. Each sample at Addgene has four layers of location which can...pages! Resources Avaliable on the Addgene Blog Read more Inside Addgene Posts! Learn all about Plasmids ...Every plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the... a microcentrifuge tube. Once a sample enters Addgene’s lab, a series of events is triggered. Each step...fluctuates throughout the year (see graph below). Addgene scientists inspect each sample and make sure they...takes over and prepares the sample for entry into Addgene’s repository. For long-term storage in our facilities... -
Caltech Systemic Capsids
TypeCollection...Abbreviations: AAV, adeno-associated virus; GRE, gene regulatory element; NHP, non-human primate. Image...Ready-to-use AAV available from Addgene's viral service. Engineered systemic constructs from Viviana ...As part of our Viral Vector Packaging Service , Addgene is distributing ready-to-use viral preparations...depth descriptions of each serotype, check out Addgene's blog on Systemic Capsids . To review performance...data of each of these serotypes, please refer to Addgene’s Data Hub and the Caltech CLOVER Center’s repository...produced with the pUCmini-iCAP-PHP.eB plasmid (Addgene #103005) . Note on PHP.eB tropism and specific ... BiSSTe4 dTomato/nlsdTomato Control Fishell Chemogenetics 44361 pAAV-hSyn-DIO-hM3D(Gq)-mCherry Syn Activator...