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We narrowed to 68 results for: lentiviral production

Showing: 21 - 40 of 68 results

  1. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...destination vector creates two products: the desires expression clone and a byproduct containing the ccdB gene...Gateway cloning to generate our desired constructs: lentiviral expression of the human KRAS gene in mammalian...sites.  Method A: recombination of an attB-PCR product or plasmid with an attP donor vector. In this case...entry clone: recombination of an attB-flanked PCR product with an attP-containing donor vector.   Method...level, and experimental purpose. For mammalian lentiviral expression, we could use a vector like pLenti...gene. Since the ccdB product is toxic to the cell, Gateway cloning efficiency can reach >99%.   STEP... tube, speeding up the cloning of the attB-PCR products directly into destination vectors. The cloning...

  2. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    Published
    Jan. 6, 2017, 3:56 p.m.
    ...work. After all, as we like to sayat Addgene, productivity is infectious. Curious which viruses researchers...lentiCas9-Blast Most of Addgene’s lentiviral inventory is based on CRISPR tools, though we do provide a...requested lentivirus and AAV of 2016. The top viruses of 2016 were (drumroll please)... Top Lentivirus: lentiCas9... Cas9-expressing lentiviruses will no longer be carried over to the gRNA lentiviruses. This means that...lentiCas9-blast lentivirus from Addgene, you could use it in conjunction with gRNA lentiviruses easily made...Viral Service and began delivering ready-to-use lentivirus and adeno-associated virus (AAV) to scientists... 2016, we expanded our viral inventory to 25 lentiviruses and 25 AAVs. These viruses have been distributed...

  3. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    Published
    Sept. 14, 2021, 1:15 p.m.
    ... Viral vectors are prone to recombination In lentiviral vectors, recombination often occurs between the...full length plasmid.   Figure 1: The full lentiviral plasmid is expected to be >12 kb. However if ...small colonies. Bacteria containing the recombined product tend to grow faster, so we recommend picking the...noticed that in Stbl3 bacterial cultures with lentiviral vectors, the flat white colonies contained the... Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3. 3 Biotech... (2007) Improved recombinational stability of lentiviral expression vectors using reduced-genomeEscherichia...

  4. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ...starting material, the ligation products are a diverse pool of lentiviral plasmids. Multiple rounds of digestion...digestion and ligation steps to build barcoded lentiviral plasmids containing one or more gRNAs. As each...for CombiGEM-based assembly into the CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from...from the pAWp28 storage vector to the pAWp12 lentiviral vector should retain the diversity in your gRNA...recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library." Nature biotechnology...

  5. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...Resources on addgene.org Addgene's Lentiviral Guide Most popular lentiviral plasmids γ-Retrovirus Guide References... the lentiviruses (like HIV-1) — that have been developed into lentiviral vectors. Lentiviruses are sometimes... 1: Gamma-retrovirus genome (left) and an HIV lentiviral genome (right.)  HIV seems an unlikely candidate...the full wild-type viral genome can direct the production, packaging, and release of progeny viral particles...Addgenie Rebecca Hutcheson for her assistance with lentiviral replication.  Resources and References More ...PMCID: PMC372876 Dong, W., & Kantor, B. (2021). Lentiviral Vectors for Delivery of Gene-Editing Systems ...and viral vectors. Key to the wild-type virus’ reproductive cycle is the fact that the virus must have its...

  6. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...efficiency. Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector...more appropriate? If you already have an end product in mind, steps 1 and 2 will generally be straightforward...plasmids provide the necessary components to make lentiviral particles. Stable, tunable expression of Cas9...: Traffic Light Reporter System: Can evaluate lentiviral component delivery as well as genome repair by...Difficulty Very easy Very hard Viral delivery   Lentivirus Retrovirus AAV Cells Dividing or nondividing...

  7. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...cells (the PCR product should contain the wild-type BRAF sequence). Then, mix the PCR products and use temperature...the middle of the PCR product. If the mismatch is in the middle of the PCR product, you would expect two...digestion products (that would co-migrate on a gel) instead of two different sized digestion products (which...middle of the PCR product, you would still be able to identify the cleavage product. 3. The Surveyor kit...what confirms it’s a real digestion product and not another PCR product. Figure 5: gRNA-targeted...GFP-edited amplification products (BRAF-GFP duplex) and from GFP-edited amplification products (GFP-GFP duplex)...with Lentivirus Isolating a Monoclonal Cell Population by Limiting Dilution Lentivirus Production Websites...

  8. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ...by targeting six different genes whose protein products have distinct localizations. Figure 2: ... The Human GlycoGene CRISPR Library is a new lentiviral CRISPR library that targets human glycosyltransferase...inhibition pooled library is now available as lentiviral preps in addition to the pooled library format...library inhibits over 18,000 human genes. Find the lentiviral preps at Addgene hSyn1-SIO-stGtACR1-FusionRed...

  9. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...within the lentiviral 3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral integration...license. In this iteration, the size of the lentiviral insert was minimized to reduce recombination ...III transcript, the sgRNAs were encoded on the lentiviral minus strand in the 3’ LTR. These optimizations...then splicing their mRNAs, the functional gene product is reconstituted within the target cells (Figure...

  10. Addgene Protocols Go Viral: AAV Purification and Titration

    Type
    Blog Post
    Published
    June 13, 2019, 12:19 p.m.
    ...2016, Addgene has shipped thousands of AAV and lentiviral preps to scientists around the world. In the ...scientists have optimized many steps in the viral production and quality control process to improve both the...

  11. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...considered safe for clinical applications.     Lentiviral: Lentiviral vectors infect dividing and nondividing...Excisable Cre-lox lentiviral vectors: This approach is the same as using a standard lentiviral vector, but ...available from Addgene. Another drawback is that lentiviral vectors insert in different genetic locations...could be affected by the location of insertion. Lentiviral insertion can also disrupt the expression of ...phenotype in infected cells. Like retroviruses, lentiviral delivery is not considered safe for clinical ... implement and doesn’t require time-consuming production of virus. In theory this is a non-integrating...plasmids if you’re interested in Tet-inducible lentiviral expression of OSKM reprogramming factors: ...

  12. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    Published
    Sept. 17, 2019, 12:40 p.m.
    ...neuroinflammation and anxiety by modulating the production of the pro-inflammatory cytokine interleukin ...closed arm (safe). Mice with higher IL-1 beta production spent more time in the open arm and were less...depression. The researchers found that injecting lentiviral vectors expressing BDNF in the hippocampus causes...Chandrasekar, and Jean-Luc Dreyer. "Selective lentiviral-mediated suppression of microRNA124a in the hippocampus...floating. In the same paper, the authors used lentivirus to introduce an shRNA to downregulate expression...

  13. Podcast: Interview with Addgene's Founders

    Type
    Blog Post
    Published
    April 13, 2018, 12:51 p.m.
    ...Addgene would be successful and the importance of lentiviral vectors in ensuring that success Chapter 6 - ... Eddy Page! Edward B. Page is a Digital Media Production Specialist at Addgene. He is the audio engineer...

  14. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ... MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with just the...to create a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed... restriction site and a linker region. The PCR product is then ligated into the MCS/SacI site in pUCXKT...occur in this system if a small contaminating PCR product formed from primer dimers and containing the missing... travels along the mRNA to produce the protein product. The system, cleverly termed TRICK, for Translation...collection of plasmids comprising the multiple lentiviral expression (MuLE) system from the laboratory ... combinations of genetic elements in a single lentiviral vector provides a powerful tool to modify mammalian...

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

    Type
    Blog Post
    Published
    May 10, 2016, 2:30 p.m.
    ...this approach could permit some viral protein production, so researchers are also interested in precisely... with HIV-1. For two HIV-1 strains, Cas9/gRNA lentiviral expression significantly reduced the HIV-1 copy...only after stable expression of Cas9/gRNA. Viral production is higher in this model than in latent HIV-1 ...PubMed Central PMCID: PMC4778041. Find CRISPR and lentiviral packaging plasmids used in this publication at...people worldwide are currently infected with the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral...

  16. 2023 and 2024: Reflections and Looking Ahead

    Type
    Blog Post
    Published
    Jan. 23, 2024, 2:15 p.m.
    ...comprehensive transcription factor library, in lentiviral vectors. At 3500 plasmids, this represents the...conversation and collaboration.  Wishing you a healthy, productive, and thriving 2024, mirroring our own aspirations...

  17. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...components to reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in...cell therapy more accessible by simplifying the production process and could even enable the creation of...cytoplasm and can be fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged... to target multiple delivery systems, such as lentivirus and MMLV gag, to specific cell types. It can ...

  18. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...incapable of producing infectious virus (similar to lentiviral vectors) because they do not encode all of the...minigenome, thus modeling genome replication. These products can be detected by Northern blot analysis. In ...Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV Guide  ...

  19. Reagent Repositories Are Speeding up Science During the Pandemic

    Type
    Blog Post
    Published
    Aug. 4, 2020, 1:15 p.m.
    ...useful to the study of SARS-CoV-2 are the lentiviral production and packaging vectors VSV-G and psPAX2, ...to scientists around the world. This scale of production and distribution is possible due to the dedicated...

  20. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ...transfection, and does not necessarily indicate the production of virus. Alternatively, if the virus encodes...packaging cells. Another method of ensuring viral production is to perform PCR against components of the viral...with, there’s a lot that goes into virus design, production, and transduction. Once you’ve harvested, it’...out Our Primer for Beginners! Learn All about Lentiviral Vectors Additional Resources on the Addgene....
Showing: 21 - 40 of 68 results