We narrowed to 1,147 results for: Gene

...Many neuroscience experiments that require gene expression in a specific cell type rely on transgenic... (recognized by Flp), to either flip or excise a gene that is located between them. Early recombinase-...Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678. https://doi.org/10.1089/hum.2019.277...Peripheral Nervous System and Relevance to Human Gene Therapy. Front Mol Neurosci 13. https://doi.org/...end of the transgene. Since loxp and lox2272 can not recombine, this ensures that the transgene is no longer...flank a transgene (homologous pairs) or used different scrambled sequences to flank the transgenes (non-... express Cre or Flp. Here at Addgene we quantify the amount of transgene recombination for every lot of...

...function better. In that case, gene shuffling might be a good option. In gene shuffling, you chop up your... this goal, including error-prone PCR, degenerate primers, gene shuffling, and structure-guided directed...mutate specific regions of your antibody. Gene shuffling Perhaps you like bits and pieces of an antibody but...sequences. Degenerate primers Using degenerate primers is similar to error-prone PCR. Degenerate primers ...resources on Addgene.org IPI Antibody Collection Antibody Guide Addgene Antibody Collection Addgene Antibody...the same general cyclical process (Figure 2). The differences typically come in when generating the library... 2: General process of antibody directed evolution. Created with BioRender.com.   Generating mutant...

...neurotransmission. But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002) and many ...2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters 520... exhaustive refinement, it was not possible to generate an optimal Rluc8-Gα construct for two of the 16...s0014-5793(02)02775-8 Additional resources on the Addgene blog Browse all of our fluorescent protein blog...Find other fluorescent biosensors Resources on Addgene.org Browse the catalog of fluorescent protein plasmids...

...experiment. PITChing MMEJ as an Alternative Route for Gene Editing Michohomology-mediated end joining (MMEJ...template to increase efficiency of insertion and gene replacement. Care to share your favorite CRISPR ...Blugene and I represented Addgene at the recent Keystone meeting on Precision Genome Editing with Programmable...hopes to evaluate genome editing pipelines and generate a common lexicon for genome editing studies. These...desired edit, and I want to make sure that I (and Addgene) don’t forget this in the future. Here’s a list... to keep current with the latest and greatest! Addgene resources 3 Tips to Improve HDR Efficiency for ...

...with genes central to the replication process removed. This means the vector can deliver a genetic payload...vectors to reproduce when experimentally necessary, genes for the required proteins are packaged in separate... More resources from Addgene The AAV Data Hub Addgene's Viral Services Addgene's Viral Protocols...viruses bud out, instead of lysing, but they do generally still kill the infected cell.) This is in contrast...labs. Viral vectors are viruses that have been genetically modified to either limit or completely eliminate...10.1007/978-3-319-07758-1_2 More resources on the Addgene blog Viral Vectors 101: Viral Applications Viral...

...sequence of a thermostable DNA ligase-encoding gene." Gene 109.1 (1991): 1-11. PubMed PMID: 1756968. 7. ...nick-closing activity of bacteriophage T4 DNA ligase." Gene 76.2 (1989): 245-254. PubMed PMID: 2753355. 12. ...blood-borne bacterial pathogens by use of a novel 16S rRNA gene PCR-ligase detection reaction-capillary electrophoresis...restriction cloning), adaptor ligation for next generation sequencing (NGS) library construction, and circularization...fragments end-to-end without the exo/polymerase generation of annealed complementary regions. Why use a... does fidelity mean for a ligase? DNA ligases generally prefer fully Watson-Crick base-paired dsDNA substrates...as a correct base pair. DNA ligases have been generally found to have a higher discrimination at the upstream...

...horizontal gene transfer, which occurs when bacteria of different evolutionary lineages share genes that can...bacteria that occurs when bacterial cells regulate gene expression in response to changes in cell density... the ASM General Meeting (now Microbe) in 2013 in Denver, CO. While I had been to the General Meeting ... were very valuable when shared with the next generation of scientists. The connection between mentors... with experience mentoring in STEM, including Addgene’s Joanne Kamens. Expanding Binning Singletons to...Binning Singletons, Inc. Additional resources on the Addgene blog Browse our blog posts on scientific conferences...

...description of the VGS process and use cases in Human Gene Therapy. Existing methods for AAV quality control...Adeno-Associated Viral Genomes in Academic Settings. Human Gene Therapy Methods 30:195–205 . https://doi.org/10.1089...and tips Resources on Addgene.org Find AAVs at Addgene Learn more about Addgene's viral service Find protocols...we partnered with Seqwell to establish a next-generation sequencing (NGS) QC platform that allows us to...methods, we showed that the DNA extract is a heterogeneous mix of single-stranded and double-stranded species...interest.   While the VGS process isn’t new to Addgene, we are excited to share the procedures and Python...Vector Preparations by Single-stranded Virus Next-generation Sequencing. Molecular Therapy - Nucleic Acids...

... vector that is better suited for analyzing the gene’s function. The process is shown graphically in the...figure, in which we add EcoRI and NotI sites to Your Gene of Interest (YGOI) for ligation into a recipient..., you should use a DNA analysis tool, such as Addgene’s Sequence Analyzer, to allow you to identify which...restriction site to improve cutting efficiency. You can generally add any 6 bases, but you should ensure that the...with 18bp of homology to the ORF). We now need to generate the reverse-complement of this sequence so that...that we can successfully amplify the ORF. You can generate the reverse-complement using existing software...the final product. Additional Resources on the Addgene Blog Learn About Other Cloning Techniques Catch...

...also allows for screening scFv generated from the B cell antibody genes from diseased or vaccinated humans...   Some phage display libraries screen antibody genes from B cells of immunized animals, but it's possible....1791  Deal CE, Balazs AB (2015) Vectored antibody gene delivery for the prevention or treatment of HIV ...also allows for generations of scFvs against two antigens that are impossible to generate scFvs against ...probe, but is easier and cheaper to generate since it’s genetically encoded.   Translational applications... of a tagged protein. Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies...-linker-VH and VH-linker-VL configurations can generate functional scFvs; however, some individual scFvs...

...amplification kit. In this case, regions of gene S and Orf1ab gene are amplified. RPA reactions also contain...amplification by primers that target the E and the N2 gene of SARS-CoV-2. If the Cas12a-sgRNA complex binds...like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas based COVID-19 testing ...protocol for Cas13-based detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA ...of temperatures varying between 10-50°C. Visit Addgene's COVID-19 Plasmids and Resources Many thanks ...Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic Material. Cold Spring Harbor Laboratory. https:...20detection%20(updated).pdf Additional resources on the Addgene blog Read more about SHERLOCK and DETECTR Learn...

...successfully treated with a CRISPR gene editing therapy personalized to his genetic mutation. “Baby KJ” was diagnosed...(2025). Patient-Specific In Vivo Gene Editing to Treat a Rare Genetic Disease. New England Journal of ...born with a C→T mutation on one allele of his CPS1 gene, which could be repaired with an A→G edit on the...administered the first dose of his personalized genetic medicine (Musunuru et al., 2025). Much has been...collaborative scientific effort. As we like to say at Addgene: sharing speeds science. Here, we’d like to highlight...in cellular models of KJ’s CPS1 mutation. Every genetic target is slightly different, so more options mean...Kleinstiver’s lab engineered new variants of S. pyogenes Cas9. While the wild-type enzyme requires a PAM...

...Gibson assembly, imagine that you want to insert your gene of interest into a vector with a large tag at the...by using Gibson assembly, you can insert both the gene of interest and the tag sequences into the vector...CATCH enables one-step targeted cloning of large gene clusters. Nat Commun. 2015; Sep 1;6:8101. PubMed...the J. Craig Venter Institute in 2009. Here at Addgene we added this option to our drop down menu of common...which chews back the 5’ ends of the fragment, generating long overhangs which allows the single stranded...advantage is that this process makes it easy to generate wild type and mutant constructs at the same time...primer melting temperature greater than 48℃. Both Snapgene and NEB have tools that help you design primers...

...may be the hot new way to manipulate gene expression, but other gene manipulation systems remain valuable...important ways to spatially and temporally control gene expression, especially in in vivo models, and new...at least six generations. In addition to the neuronal pThy1-Brainbow constructs, Addgene also has two ...Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0, 1.1, 2.1 plasmids...coupling of Cre-lox with FLP-FRT recombination to generate additional recombination events, and the synthesis...and sea anemone mKate2.) They then successfully generated custom antibodies to each of these proteins and...analysis. To achieve even cell labeling, they generated farnesylated derivatives of the fluorescent proteins...

... provide a highly versatile means of delivering genes to many different cell types with minimal immunological...effects. AAVs are used in experiments targeting genes to specific cell types in the brain, mapping neuronal...Resources at Addgene.org Addgene's Viral Service Find Retrograde AAV Browse Optogenetics Plasmids ... to our viral vector pages on the Addgene website and on the Addgene blog to get some ideas for future...Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering Primer on Optogenetics Resources... Addgene recently attended the Society for Neuroscience 2017 conference in Washington DC (#SfN17). This...newbies might want to get into the field. Find Optogenetics Plasmids for Use in Neuroscience Research Neuroscience...

...were compared head-to-head for gene expression changes of non-target genes and RNA-seq of a nuclear (MALAT1...among the first batch of antibodies we produced at Addgene and, as you might suspect, it has been one of our... need a second GFP antibody?  Similarities:  Generated against full length GFP and converted to recombinant...always best to have options.      Fig. 2: Addgene’s western blot for mEGFP-HRas (from Plasmid 18662... [N86/8R] page for more details. Image credit: Addgene.   Find anti-GFP [N86/8R] here!   Andrews NP...

...Pancreatic and duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin-induced...lentiviral vectors can be used for gene delivery Resources on Addgene.org Visit our Stem Cell Pages Find...from the patient in vitro then correct the damaged gene in the resulting iPSCs before differentiating the...PMC3987909. 6. Ban, H., et al., Efficient generation of transgene-free human induced pluripotent stem cells...Mukherjee. The ultimate goal in the field of regenerative medicine is to replace lost or damaged cells...cell is converted to a different cell type for regeneration and repair and situations where one process ...plasmids used for these methods can be found on Addgene’s stem cell page. On this page, you can also find...

...dCas9 and microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP system...Fig. 3: Development of a cleavage-free dCas9-based gene editor using microbial SSAPs. Image from Wang, et...reduces the need to generate “single-use” transgenic lines. Instead, researchers can generate and maintain transgenic...each protein of interest and, for in vivo work, generate a dedicated transgenic line. Using zebrafish as... the liver.    Fig. 2: Summary of the generational evolution of the AAV capsids (left) and transduction...

...enzyme cloning uses short sticky ends to insert genes into plasmid backbones, base pairing between these...Schematic of SLIC. To start the SLIC cloning process, a gene of interest is PCR amplified to add 5′ and 3′ homology... LIC received an important update, courtesy of Addgene depositor Stephen Elledge. His new method, named...flexible. For one thing, those 5′ overhangs can be generated in several ways. One option, imprecise T4 digestion...work directly on PCR products, with 5′ overhangs generated either by annealing a mixture of products or from...regions from the target vector. 5′ overhangs can be generated by T4 polymerase (left branch); by annealing two...concentrations of insert to succeed. 5′ overhangs are generated in the linearized vector by T4 digestion; then...

...observed in mammalian cells is to tag an endogenous gene. By imaging cells (or tissues) that produce an endogenous...fluorescent proteins or using endogenous tagged genes) can be used to verify performance under different... develops genetically encoded FRET based biosensors.     Additional Resources on the Addgene Blog Which...mVenus as a reference protein is available from Addgene. Another way to dodge cellular variation observed...Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes. You can follow him... Protein Biosensors Additional Resources on Addgene.org Find Empty Backbones with Fluorescent Tags Find...