We narrowed to 1,145 results for: Gene
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TypeCollection...Adeno-associated virus (AAV) vectors are safe and useful gene delivery tools that effectively transduce many mammalian...Packaging Service overview. Function Chemogenetics DREADDs, PSAMs Optogenetics Channelrhodopsins, Halorhodopsins...Ready-to-use adeno-associated virus (AAV) available from Addgene's viral vectory packaging service. Browse constructs...preps are purified by density centrifugation. Addgene’s comprehensive high-quality quality control process...shipment. If you can't find the prep you need, try Addgene's AAV Packaged on Request service. You can choose...
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CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog Post... (Shmakov et al., 2015). They were using Cas1, a gene commonly associated with CRISPR arrays and involved...analysis, they identified 53 potential candidate genes that fell into 3 categories based on the architecture...expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although the...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Check Out Our CRISPR Featured...27, 2020. CRISPR, and specifically Cas9 from S. pyogenes (SpCas9), is truly an exceptional genome engineering... PMC5127784. Find plasmids from this paper at Addgene. 2. East-Seletsky, Alexandra, et al. "Two distinct...PMID: 27669025. Find plasmids from this paper at Addgene. 3. Gootenberg, Jonathan S., et al. "Nucleic ... -
MXS Chaining
TypeBlog Post...inducible gene-expression sequences, and more. By combining these components, one can generate constructs...localization, as well as cellular processes like gene expression and cell migration (Sladitschek and Neveu... might not, however, be appropriate for cloning genes from organisms with high CpG dinucleotide representation...to combine the functions of different genetic elements to generate non-natural tools such as novel biological...Resources on Addgene.org Browse our Fluorescent Protein Collection Find Plasmid Kits at Addgene Find Plasmid...nutshell, is the systematic combination of different genetic sequences into plasmid DNA. In high throughput ...techniques, although the specific sequences of the genetic elements may differ (e.g., a set of various mammalian... -
Neuronal labeling with Spaghetti Monster
TypeBlog Post...nucleus AAVs CREATEd for gene delivery to the CNS and PNS Resources on Addgene.org Find the Spaghetti Monster...with the development of methodologies to deliver genes to the CNS have recently made it much easier to ..., and the long-term expression of AAV delivered genes. AAV serotypes have been extensively studied in ...essential for neuroscience research. The use of genetic tools for visualizing and perturbing circuits together...Adeno-associated virus (AAV) has been widely used to deliver genetic tools to the brain because of its low cytotoxicity...connections to that population. Limitations of common genetically encoded neuronal tracers Neuronal tracers such...influenza hemagglutinin (HA), myelocytomatosis viral oncogene (myc), simian virus 5-derived epitope (V5), the... -
CRISPR 101: Making a Knock-In Cell Line
TypeBlog Post...-in mutations and gene editing Targeted genome editing events fall into two general classes: knock-in ...HDR. If your Cas9 cuts in the coding region of a gene, make sure that the PAM edit you introduce is a ...Guo, Y., Tian, Qigi, Lan, Y., et. al. An efficient gene knock-in strategy using 5’-modified dsDNA donors...translation by generating a frameshift mutation. The majority of cellular repair events will generate these types...family member which generates sticky ends upon cutting. Cas12a may promote HDR by generating ssDNA overhangs...Taking the road less traveled and generating a knock-in cell line instead of a knock-out? We’ve got you...cells, but a less frequent DNA repair mechanism in general. On average, it accounts for less than 10% of DNA... -
Cpf1: A New Tool for CRISPR Genome Editing
TypeBlog Post...directional gene transfer, analogous to traditional restriction enzyme cloning. Sticky-end mediated gene transfer...from this publication at Addgene. 2. Zetsche, Bernd, et al. "Multiplex Gene Editing by CRISPR–Cpf1 Using...26411297. Resources at Addgene Find CRISPR plasmids at Addgene Read Addgene's CRISPR Resource Guide View...Again, they were in for a surprise! Whereas Cas9 generates blunt ends 3 nt upstream of the PAM site, Cpf1...27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik, et al. Genome-wide analysis reveals...27272384 Find plasmids from this publication at Addgene. Kleinstiver, Benjamin P., et al. "Genome-wide...27347757. Find plasmids from this publication at Addgene. Makarova, Kira S., et al. "An updated evolutionary... -
A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression
TypeBlog Post...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... -
Antibodies 101: Antibody Engineering and Directed Evolution
TypeBlog Post...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... -
Fluorescent Protein Guide: Empty Backbones
TypeCollection...you to create fusion proteins with your gene of interest....protein (GFP) was cloned in 1992 ( Prasher et al., Gene, 1992 ), and since then scientists have engineered...tags for you to create fusion proteins with your gene of interest. These are organized By Color of the...Lab Lentiviral Plasmids - Fluorescently tag your gene for lentiviral expression Verkhusha Lab Plasmids...Addgene has assembled a collection of empty plasmid backbones with different fluorescent tags for you...Monomer? Background Fluorescent proteins are genetically encoded tools that are used extensively by life...proteins that result in a diverse set of colors. Addgene has assembled a collection of empty plasmid backbones... -
CRISPR Challenges: Standardization and Homology Directed Repair
TypeBlog Post...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 ... -
Viral Vectors 101: What is a Virus (and a Viral Vector)?
TypeBlog Post...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... -
Understanding GPCR Signaling Using TRUPATH
TypeBlog Post...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... -
When Fidelity Matters: A frank discussion about ligase fidelity
TypeBlog Post...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... -
Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone
TypeBlog Post...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... -
Chemogenetics Plasmids
TypeCollection...AAV preparations of many chemogenetics plasmids. ID Plasmid Description Gene/Insert Vector Type Promoter...Service: Chemogenetics AAV Blog: Chemogenetics Viral Vectors 101 Blog: Chemogenetics vs. Optogenetics Chemogenetics... more about chemogenetic resources available at Addgene. See our inventory of chemogenetic constructs,...Collections Chemogenetics Plasmids Chemogenetics Plasmid Collection You may also like... Chemogenetics Guide...biological processes. Addgene’s collection contains two main classes of chemogenetic receptors: Designer...luciferase and a light-sensing optogenetic element, making a bimodal opto-chemogenetic actuator. When the luciferase...your experiments, please see our Chemogenetics Guide . Chemogenetics Plasmids Browse, sort, or search ... -
Resolute Plasmid Collection
TypeCollection... of Gateway-compatible pDEST vectors. ID Plasmid Gene/Insert Industry PI Pooled Libraries CRISPR and overexpression...engineering forms the core of the deorphanizing engine. Genetic and proteomic interactions represent the framework.... Protein production is used for assays and to generate antibody/ high-affinity ligands. Data is embedded...integrated knowledgebase that features, among others, genetic variants and disease associations. Plasmids This...overexpression pooled libraries are useful tools for genetic screening experiments. These libraries target the...Plasmid Collection NeuroMab Plasmid Collection Addgene Blog: Gateway Cloning Content last reviewed: 12... -
Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation
TypeBlog Post...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... -
Plasmid Cloning by PCR
TypeBlog Post... 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... -
SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas
TypeBlog Post...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... -
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog Post...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...