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We narrowed to 65 results for: lentivirus expression plasmid

Showing: 41 - 60 of 65 results

  1. DAP arrays for multiplex gene editing

    Type
    Blog Post
    Published
    July 7, 2022, 1:15 p.m.
    ...promoter, which is used to drive the expression of the array. After expression, the individual gRNAs are released...demonstrated that the tRNA-gRNA array can drive the expression of itself, without using any additional promoter...deliveries via adeno-associated virus (AAV) and lentivirus, demonstrating simultaneous editing of genes ..., that rely on an upstream lengthy promoter to express the gRNA array, DAP arrays use tRNA itself, as ...tRNA-gRNA arrays, which is scalable and efficient in plasmid and viral vectors. In applications when repetitive...manual and online design examples of using Addgene plasmids hCtRNA_FT (#186715) and hCtRNA_VT (#186716).   ...resources on the Addgene blog CRISPR 101: Multiplex expression of gRNAs Four Base Editing Reporters to Monitor...

  2. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    Published
    April 13, 2023, 1:15 p.m.
    ...applications Gene expression control Researchers are always wanting to manipulate gene expression – turn it up...interest, turning expression off permanently. Viral vectors can also be used to express proteins. Viral... the genome of the target cell or transiently expressed. It’s important to note that for all the applications...is more than one type of virus to choose from (lentivirus, retrovirus, AAV, etc.). When you’ve decided ...sequence for degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily...Viral expression cassettes typically contain a promoter (inducible or constitutive), your gene of interest...and often a selectable marker. This vector can express your construct in target cells transiently or constitutively...

  3. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    Published
    Feb. 20, 2014, 4:57 p.m.
    ... smaller size of the plasmids. The Zhang lab, on the other hand, uses a 1 plasmid, single infection system...technology.” GeCKO library for genomic screens Using lentivirus as the delivery vehicle, Neville Sanjana, also... thousands of targets at once. That’s what our plasmids allow.” Wang’s library is designed to target a...have previously been stably transfected for Cas9 expression. The Yusa lab has used their library to screen...

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

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...provide synthesized SARS-CoV-2 expression plasmids available for many expression systems: bacterial, yeast,...small collection of plasmids for coronavirus research, including a handful of plasmids that have been in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate... pseudotyping lentivirus with the SARS-CoV-2 spike protein. They deposited the plasmids used for this ...spike protein to improve expression and pseudotyping efficiency. This plasmid was later used by another...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from...

  5. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ... to co-express them from one promoter. Specificity The ability of AAV to drive gene expression in specific...cell-specific gene expression and viral infection. Cre or Flp dependent switches control gene expression via switches...transgene expression, since you don’t have to wait for the host to replicate the DNA before expression. Now...non-enveloped icosahedral capsid, that can be used to express genes of interest in cell and animal models. AAVs...laboratory experiments ranging from transient gene expression in a particular cell type to CRISPR genome editing...genes, you can coinfect with multiple viruses to express genes or gene sets over 4.7 kb. Co-infection rates.... On the other hand, if you’re interested in expressing multiple small genes, you can design single rAAV...

  6. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...transgene expression. Once packaged in AAV, this pre-recombined DNA could lead to transgene expression in recombinase-negative...sufficient transgene expression but limits the likelihood of recombinase-independent expression. Next-generation...Ready-to-Use AAV Find Ready-to-Use Lentivirus Browse Viral Plasmids ...serotype. Additionally, we check viral transgene expression in transduced cells whenever possible. This post... tube is mislabeled or someone grabs the wrong plasmid? What if bacteria snuck into the prep during the... human error, fragments of DNA from the helper plasmids or the cell genome can be packaged inside the ...) are aligned to the reference sequence of the plasmid used to create the viral prep. In a clean viral...

  7. Viral Vectors 101: Biosensors

    Type
    Blog Post
    Published
    July 6, 2023, 1 p.m.
    ...analyte), and accumulation of ATP activated GFP expression (receptor), then the transducer could be a microscope...genetically encoded, and luckily, we have pre-designed plasmids for these systems! Ion sensors One common biosensor...retrovirus, can either permanently integrate and/or express long term (up to several years!). Biosensors tend...biosensor to specific tissues or cell (sub)types for expression and downstream monitoring. Nervous about making...ready to deliver your sensor, whether you choose lentivirus, gamma retrovirus, or AAV as your vehicle, we...we’ve got you covered with packaging plasmids! References and Resources References  Bhalla N., Jolly ... as stable systems, rather than transiently overexpressing at the time of use. Another great viral perk...

  8. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ...precisely control where and when certain genes are expressed to investigate their role in a specific behavior...Which viral vector should be used? Modifying gene expression Generally speaking, one may want to achieve either...application.  Genetic manipulation can also reduce expression levels of certain genes. A popular method uses...allows scientists to draw conclusions due to gene expression in a specific brain area, avoiding side effects...targeting the gene is cloned into a viral shRNA expression vector and the vector particles are delivered...delivered into the brain region of interest. AAV or lentivirus?  Both AAV and lentiviral vectors are used for... a tight spatial and temporal control of gene expression manipulation. They are injected using a stereotactic...

  9. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    Published
    Feb. 24, 2015, 7:50 p.m.
    ...tetraloop and stem-loop 2; 3. The MS2-P65-HSF1 plasmid which expresses the activation helper protein. SAM can ...Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation in mind...that will enable even more fine-tuning of gene expression levels and on software for analyzing the data...you are equipped and authorized to make and use lentivirus. Also, note that due to the size of the backbone...

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

    Type
    Blog Post
    Published
    Aug. 18, 2023, 4:15 p.m.
    ...required proteins are packaged in separate plasmids and expressed via cells cultured with the vectors. This...after the viruses they’re derived from: AAVs, lentivirus, gamma retrovirus, and adenovirus. And now that...

  11. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. .... Today, it's time for another edition of Hot Plasmids!  Here's what you'll find in this post:  New...., 2023 under CC-BY 4.0 License. Find PE6 plasmids here!  Doman, J.L., Pandey, S., et al. (2023).... to target multiple delivery systems, such as lentivirus and MMLV gag, to specific cell types. It can ...therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D., et al. (2023). Cell type-specific...pulse-chase experiments, and more. Find LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID...LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments open up new possibilities...

  12. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...packaging plasmids: the pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing...HEK293T cells, expressing Ad E1A and E1B, can be transfected with your transfer plasmid, carrying your... interest, and the two helper plasmids, commonly called “triple plasmid transfection” (Figure 1).    ...production process.  Helper-free production Unlike lentivirus, AAV does not possess the ability to replicate...repeats (ITR). You should always have a high-quality plasmid standard and rAAV reference material of a known...before undergoing titering, to remove residual plasmid DNA carried over from the production process. For...Walther, N., & Maurer, A. C. (2023). Transgene Expression in Cultured Cells Using Unpurified Recombinant...

  13. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...Endogenous biotin ligases Expression system Stable cell line expressing 1) FLAG-biotin-tagged dCas9,...requires the creation of a stable cell line that co-expresses three components: dCas9 with an added biotin ...interest  When these three components are co-expressed, the gRNA targets the dCas9 to the loci of interest...lines will have varying levels of dCas9 and gRNA expression. Additionally, CAPTURE 3C-based sequencing requires...that is recognized by biotin ligases normally expressed in eukaryotic cells. Using this tag reduces the...CAPTURE 2.0 to be used with primary cells. Find plasmids for CAPTURE 2.0! When compared to the original... BirA, and 3) one or more gRNAs Two separate lentiviruses delivering 1) a BioTAP-tagged dCas9 and an eGFP...

  14. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ... Lentiviral Plasmids Tetracyline Inducible Plasmids Empty Backbones Bacterial Expression Systems AAV ... that made it easy to share plasmids, both for people depositing plasmids and for people requesting them...AAV Plasmids Blogs Origin of Replication What is a Plasmid? How to Design Your gRNA for CRISPR Editing... Verify Your Plasmid Using a Restriction Digest Analysis Videos What is a Plasmid? How to Design...video, however, is “What is a Plasmid?”, An animated explainer of a plasmid and its parts that helps the...them. Prior to this, many plasmids were available, but they often weren’t easily accessible, due to a ...barriers, allowing people to access a wide catalog of plasmids from all over the world. This idea, that available...

  15. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...immunogenicity and its transient expression. LV results in long term expression in dividing and non-dividing...versatile tool for gene expression in mammals Resources at Addgene.org Find AAV plasmids Find ready-to-use...viral vectors capable of providing long-term gene expression in retinal cells - AAV - were becoming available...other viral vectors such as adenovirus (AdV) and lentivirus (LV) are known to transduce retinal cells, AAV...still lead to long-term (>10 years) transgene expression in non-dividing cells because they remain in ...engineering of new capsid variants, designing expression cassettes to regulate and optimize transgene ...

  16. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    Published
    May 10, 2016, 2:30 p.m.
    ...people worldwide are currently infected with the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral...HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can produce HIV-1 toxic proteins... gRNAs to target the HIV-1 5’ and 3’ LTRs and expressed them along with Cas9 in the T-cell line 2D10. ...of the LTRs joined together. Clonal Cas9/gRNA-expressing cell populations were also immune to HIV-1 reinfection...cycle progression, or apoptosis with Cas9/gRNA expression. A pooled analysis found no evidence of Cas9 .... For two HIV-1 strains, Cas9/gRNA lentiviral expression significantly reduced the HIV-1 copy number, ...preventing re-infection would require sustained expression of Cas9/gRNAs, which could result in higher rates...

  17. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ... DNA compaction, an enhancer can increase the expression of a gene that is millions of base pairs away...Putting it all together, we get the following expression for step 1: (H3K4me1 + H3K27ac) – H3K4me3 The...enhancers with CRISPR Armed with the genome math expression for NANOG enhancers, we’re ready for step 2: ...cells. Remove this pillar and the ESC dies. Order Lentivirus from Addgene If you’re interested in designing...Genome Edit Resources on Addgene.org Find CRISPR Plasmids for Your Research Check out Our CRISPR Guide Find...

  18. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...conditional gene expression, multiplexed CRISPR, and chromosomal integration of larger expression cassettes ...chromosomal integration and gene expression, the system includes a toolkit to express recombinant proteins within...comparisons between CRISPR components expressed from different plasmid backbones. They noted that backbone...or distance in the placement of plasmid elements can impact expression levels and consequently convolute... tools can be used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even...from individual transcription units (TUs) used to express your favorite gene-of-interest, all the way to ...with E. coli and/or cell-free systems. Protein expression in yeast MoClo is also for eukaryotes! The technology...

  19. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ...to All Things Lentivirus Resources on Addgene.org Check out this guide to pooled plasmid libraries Find...control for many of the effects of gRNA and Cas9 expression, they fail to account for the effects of Cas9...lentiviral vector to cell lines that either stably expressed Cas9 or that had been lentiviral infected with...sites should control for the effects of guide expression and dsDNA breaks. Figure 2: Non-targeting...

  20. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...amazing plasmid technologies developed by our community of depositors. With over 40,000 plasmids avaliable...amazing new plasmid tools that have come through our doors in 2015. Do you have a favorite new plasmid tool ... mediated genome modification and gene expression. The plasmids associated with this article can be obtained...system plasmids are available through Addgene and are already very popular. Use these plasmids to increase... of the integrases is supplied on its own expression plasmid or in various combinations, while the cognate...bacterial expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT...Fortunately, a collection of plasmids comprising the multiple lentiviral expression (MuLE) system from the ...
Showing: 41 - 60 of 65 results