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We narrowed to 1,146 results for: Gene

Showing: 721 - 740 of 1146 results

  1. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...situ is the knock-in of fluorescent tags into the gene locus of interest. An advantage of this alternative...studied a synaptic protein that is linked to neurodegeneration. The level of this protein is decreased in...PMCID: PMC4121734. Additional Resources on the Addgene Blog Fluorescent microscopy techniques - Which... tagging strategies Additional Resources on Addgene.org Fluorescent protein plasmids Use FRET to study...protein-protein interactions Deposit plasmids with Addgene! ...

  2. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    ...repair of Cas9-generated breaks is useful if the intent is to make a knockout in your gene of interest, ..., as it is prone to generating indel errors. Indel errors generated in the course of repair by NHEJ are...are typically small (1-10 bp) but extremely heterogeneous. There is consequently about a two-thirds chance...Of some importance, the deletion can be less heterogeneous when constrained by sequence identities in flanking...overhangs is available. Cas9 can also be altered to generate a targeted single strand break; when two such ...  Dale Ramsden is a member of the curriculum in Genetics and Molecular Biology, the Dept. of Biochemistry... Break Diminishes Its Mutagenic Potential. PLoS Genet 5:e1000683 . https://doi.org/10.1371/journal.pgen...

  3. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ...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...

  4. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...Chemical Modifications of CRISPR RNAs to improve Gene-Editing Activity and Specificity. JACS 144, 12584...resources on addgene.org  View Addgene's collection of empty gRNA vectors View Addgene's CRISPR resources...effectively serving their function of guiding Cas9 to generate a targeted break. These advances in gRNA technology...or immunity itself is being studied. Check out Addgene’s validated gRNA repository to find pre-tested gRNA...photocleavable gRNAs. Photocleavable guides are generated by introduction of a single photocleavable 2-nitrobenzyl...lentiviral plasmids Additional resources on the Addgene Blog  How to design your CRISPR gRNA CRISPR 101...

  5. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog 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...

  6. Transferable Skills Guide: Public Speaking

    Type
    Blog Post
    ...You realize, much too late, that you labeled your gene of interest incorrectly on slides 8-16. Your notes...remember related to this paper and/or biology in general until I reached Fig. 6, Panel L, and hope nobody...breath–you got this!  Additional resources on the Addgene blog Transferable Skills Guide: Time Management...

  7. RNA Extraction Without A Kit

    Type
    Blog Post
    ...downstream application such as quantitative PCR to assess gene expression. There are several things that researchers...everything, including your skin. Here are some general tips for working with RNA, even if you are using...10.1038/nprot.2006.83 Additional resources on the Addgene blog Read blog posts related to plasmid protocols...for purifying DNA without a kit Resources on Addgene.org Check out our molecular biology protocols Find...

  8. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ... useful as a mechanism of transient knockdown of gene expression. This system could be used in lieu of...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9 has been...Cas12’s primary difference from Cas9 is that it generates staggered breaks, also known as sticky ends. Cas12a...targeting experiments requiring single strand DNA generation, such as homology directed-repair. Cas12a is ... of DNA. This makes it particularly useful in generating transient edits of signaling molecules (RNA) ...If your genome engineering goals expand beyond generating frameshift mutations or single base pair edits...s41586-021-03886-5 Additional resources on the Addgene Blog CRISPR 101: Which Cas9 do I choose for ...

  9. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog 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...

  10. Rinehart Lab Phosphoprotein Reagents

    Type
    Collection
    ...for the incorporation of phosphoserine into your gene of interest....phosphoserine into the growing polypeptide chain to generate the new phosphorylated protein. Protein Phosphorylation...by adding this phosphorylated amino acid to the genetic code of E. coli (Park et al., 2011). Follow up ...further details in Mohler et al. (2023) and find general tips on phosphoprotein expression in rEcoli XpS...Barber et al., 2018). This phosphosite library was generated in a single mixed pool by expressing and purifying...assessments. The Mode #1 phosphosite library can be generated with either phosphoserine using SepOTSλ (Plasmid...synthesized using supD tRNA ( SupD (Plasmid #68307) ) to generate the phosphosites with serine instead of phosphoserine...

  11. Plasmids 101: Protein Expression

    Type
    Blog 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...

  12. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog 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...

  13. Antibody Guide

    Type
    Collection
    ...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...

  14. Technical Design of a Western Blot

    Type
    Blog 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 ...

  15. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog 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 -...

  16. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog 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 ...

  17. How to Write a Scientific Review Article

    Type
    Blog 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...

  18. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog 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...

  19. Hot Plasmids - November 2023

    Type
    Blog 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...

  20. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    ... preparations on downstream applications. Qiagen Gene Expression Newsletter. Issue 15/10. March 2010. ... Tech Tip (2013).  Additional resources on the Addgene blog Plasmid Preps: Different Purity, Different... Your Plasmid Purification Kit Resources on Addgene.org DNA Quantification Protocol Recovering Plasmid...
Showing: 721 - 740 of 1146 results