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We narrowed to 1,141 results for: Gene

Showing: 581 - 600 of 1141 results

  1. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...(MP6) permits quick, large-scale mutation of the gene of interest. They created a selection phage (SP)...transcription from the accessory plasmid (AP) encoding phage gene III. The accessory plasmid library contained all... a library PAM, it would induce transcription of gene III, permitting phage propagation and keeping that...evolution to generate and select for Cas9 mutants with PAM flexibility. In this system, the mutagenesis plasmid...this paper are available at Addgene.  Additional Resources on the Addgene Blog Learn about Engineered...another 72 hours of phage-assisted evolution, generating xCas9 2.0-2.6. This second round of evolution... Cas9 variants with activity on non-NGG PAMs, generating xCas9 3.0-3.13. To test xCas9 3.0-3.13, Hu et...

  2. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...22,000 protein coding genes in the human genome with fluorescent proteins. For each gene, we needed to design...way that prevents re-cutting but does not disrupt gene expression. (See section below.) For guide design... Zuckerberg Initiative. We hereby announce the general availability of new a tool for CRISPR scientists...analyses, but it also allows you to download any generated info for your own purposes, or delete the originals...at existing tools. As has been written here on Addgene, there has been a Cambrian-like explosion of software...that it can be used in the rank equation above. Generally, the greater this distance between the cut and...sequence inserted by HDR. Although this practice is generally advised, there is little detail published on how...

  3. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ...Click on the gene names in the figure below to find plasmids containing that particular gene. ...Additional Resources on the Addgene Blog Learn More about Plasmids for Endogenous Gene Tagging in Human Cells...FP properties. It is important to look out for general problems associated with fluorescent imaging such...constructs you create in your own work available to the Addgene community. Download our Markers of Subcellular ...PMCID: PMC4777285. Further Reading Microscopy U. Generfal Considerations when Using Fluorescent Proteins...Collection Get Tips for Using FRET Resources on Addgene.org Browse the Allen Institute for Cell Science ...

  4. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...guides and an NgAgo expression plasmid to edit a gene in vivo. Try NgAgo for Yourself On the other hand...were having difficulty achieving indel formation (gene disruption) with this new technology. To dig more...asked if they could detect indels (an indication of gene disruption ability) with NgAgo, one respondent out... do not necessarily represent the views of Addgene. Addgene performs Sanger sequencing on select regions...the water. On the bright side, successful indel generation and epitope knockin was achieved by a few respondents...Twitter - Tweet suggestions and tips with #NgAgo The Addgene Blog - Leave suggestions and tips in the comments...comments section below or contact blog@addgene.org if you'd like to write a more detailed post It may take...

  5. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...directional gene transfer, analogous to traditional restriction enzyme cloning. Sticky-end mediated gene transfer...adeno-associated viral vectors) are commonly used for in vivo gene delivery due to their low immunogenicity and range... the potential “sticky end” transgene insertion mediated by Cpf1-generated overhangs. Exploring the efficiency...suited for genome editing.  Find Cpf1 Plasmids at Addgene   Image attribution: Zetsche, Bernd, et ...in AT-rich genomes. Additionally, whereas Cas9 generates blunt ends 3 nt upstream of the PAM site, Cpf1...certain tissues. However, these vectors have a low genetic capacity of ~4.5 kb. At 4.1 kb, SpCas9 barely fits...27272384 Find plasmids from this publication at Addgene. 2. Kleinstiver, Benjamin P., et al. "Genome-wide...

  6. Zhang Lab's CRISPR Frequently Asked Questions

    Type
    Collection
    ...example, choosing target sites that span the knock-in gene. For making mutations, one good way is to mutate...be okay if you are introducing a selection marker gene. The homology arms should be no more than 100bp ...for my genomic PCR, but couldn't amplify the EMX1 gene using same primer you used in the Science paper ...You may also like... Zhang Lab CRISPR plasmids General CRISPR Plasmids Dr. Le Cong, the first author of...additional PAM sequences should not be sufficient to generate off-target DSBs; there needs to be extensive homology...below are considerations for designing HR donor. Generally if off-target (non-specific) cleavage is not a...purified oligo is fine based on new data). For general transfection, you could use 1ul of 10uM oligo stock...

  7. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ...Truncated gRNAs for Regulating Gene Expression and CRISPR 101: Epigenetics and Editing the Epigenome (plus...Joining and PITCHing MMEJ as an Alternative Route for Gene Editing Day 9: Precision editing: Single Base Editing...Since the start of the CRISPR revolution, Addgene has distributed over 100,000 CRISPR plasmids. But that...flame for its adenine base editing capabilities. Epigenetics: CRISPR has made it possible to design programmable... blue flame plasmids covering three different epigenetic effectors. The diversity of tools is especially...plus bonus podcast with Addgene Advisory Board member Alex Chavez) Day 11: Pooled libraries: Genome-Wide...

  8. SunTag and Fluorescent Imaging

    Type
    Blog Post
    ...used to improve CRISPR-based activation of target genes, but we’ll save that application for another day...protein-tagging system for signal amplification in gene expression and fluorescence imaging.” Cell 159(3...mitochondria without organelle toxicity. First generation (v1) SunTag is expressed at very low levels due...protein-by-protein basis. However, since many of Addgene’s SunTag plasmids have coveted “blue flames,” it...PMCID: PMC4252608. Additional Resources on the Addgene Blog Sign Up to Receive the Fluorescent Protein...about Small Molecule Fluorphores Resources on Addgene.org Find Fluorescent Protein Empy Backbones Visit...

  9. Fluorescent Proteins 101: Fluorescent Biosensors

    Type
    Blog Post
    ...biosensors To focus on a particular gene or protein, check out our gene-specific biosensors. These constructs...Visualization Resources on Addgene.org Find Biosensor Plasmids Find Optogenetics Plasmids Visit the Fluorescent... to the molecule it detects. This change then generates a reporter signal. Reporter signals may be electrochemical... OR enzyme activity. Small molecule biosensors Addgene has many sensors that measure the concentration..., and voltage across membranes. These sensors generally follow the same pattern described above - a sensing...fluorescent molecule   Additional Resources on the Addgene Blog Check out the Rosella Autophagy Sensor Read...

  10. Cre-ating New Methods for Site-specific Recombination in Drosophila

    Type
    Blog Post
    ...and caveats of Cre usage: Vagaries of conditional gene targeting. Schmidt-Supprian M, Rajewsky K. Nat Immunol...activity regulation. Heidmann D, Lehner CF. Dev Genes Evol. 2001 Sep;211(8-9):458-65. doi:10.1007/s004270100167...specific recombination site for each enzyme, were generated to test recombination efficiency. When expressed....1111704108. Epub 2011 Aug 9. PubMed. Find the Plasmids @Addgene: Plasmids for KD, R, B2, and B3 recombinases from...

  11. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    ...attach the gene of interest to either the N-or the C-terminus. Even if a researcher’s gene of interest...researchers as we could, and Addgene is a wonderful vehicle for us to do that.” At Addgene, we are equally excited...is available in an easily searchable format on Addgene’s Michael Davidson Fluorescent Protein Collection...collection with over 300 backbones alone,” said Addgene’s Lianna Swanson, who has been working with members...the scientific community," says Joanne Kamens, Addgene's Executive Director, "and to promote the great ...

  12. CRISPR Plasmids - RNA Editing

    Type
    Collection
    ...editing in mammalian systems. Mammalian ID Plasmid Gene/Insert PI Publication Do you have suggestions for...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource!...

  13. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog 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...

  14. MXS Chaining

    Type
    Blog 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...

  15. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog 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 ...

  16. Neuronal labeling with Spaghetti Monster

    Type
    Blog 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...

  17. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog 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...

  18. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog 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...

  19. Ginkgo Bioworks COVID-19 Collection

    Type
    Collection
    ...SARS-CoV-2 and common coronvirus plasmids based on: Gene insert (e.g. spike, envelope, nsp3, OC43, NL63, ...Additional Resources Read Addgene's Blog Synthesized by Ginkgo Bioworks, Shared by Addgene: SARS-CoV-2 Plasmids...for a summary of the Ginkgo collection at Addgene. Addgene's COVID-19 and Coronavirus Plasmids & Resources... viral proteins in several expression systems. Addgene has partnered with Ginkgo to make these plasmids...expressed or functionally validated by Ginkgo or Addgene. Plasmids Use the search bar or sort buttons to...to COVID-19 that may be of use to scientists. Addgene's Protocol for RNA Extraction Without A Kit...

  20. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog 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...
Showing: 581 - 600 of 1141 results