... Service Suggestion Form. Lentivirus As some of the most popular lentiviral vectors in the repository ... of our new Viral Service, we’re distributing lentivirus (with many CRISPR tools included among the preps...collection. We’ll provide a brief rundown of what lentiviral and AAV preps are available below, but you can... tools, we’re starting the Viral Service with lentiviral preps for the delivery of Cas9 endonuclease, ...we’ve got viruses to help you out. Of course, lentiviruses are used in experiments other than just those...those involving CRISPR and so we also have lentivirus for delivering control shRNAs, GFP, and rtTA from ...with viruses including advice on how to titer lentiviral preps, troubleshoot viral transductions, and ...

...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 ...transgene reactivation, residual expression Lentivirus Integrating High Low Efficient and Stable Genome...iPSC-transplanted mice. Cre-deletable or Tet-inducible lentiviruses address some of these concerns, but overall ...reprogramming factors driven by one promoter. While lentivirus has similar rates of reprogramming when compared...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 ...

...Sakuma et al   Multiplexed Lentiviral Expression Cassettes lentiviral transduction One vector expresses...expressing the Cpf1 multiplex system from a lentiviral vector.  Lentiviruses carry a (+) strand RNA copy of the... options for Cpf1 multiplexing: Transfection, lentivirus, and AAV Cpf1’s ability to process its own pre-crRNA... multiple expression platforms: transfection, lentiviral transduction, and AAV transduction.  In most ...Cpf1. This means Cpf1 can bind and cleave the lentiviral RNA, preventing packaging of the virus.  Reversing...of the direct repeat protects the (+) stranded lentivirus RNA from Cpf1-mediated cleavage. This reversing...

... of the AAV and lentiviral preps we have released in the last few months: Lentiviral preps for expressing...plasmids into lentiviral preps to speed up COVID-19 research even further. The following lentiviral preps are... GFP dependent Cre and FLPo And more!  New lentiviral preps SARS-CoV-2 protein expression The Krogan...2xStrep-IRES-Puro: Expression of the SARS-CoV-2 ORF3a Lentiviral preps for expressing the E protein (pLVX-EF1alpha-SARS-CoV...Dolcetto pooled library is now available as a lentiviral prep. It's a human CRISPR inhibition library ...-wide inhibition screening. Find the Dolcetto lentiviral prep here Control AAV Control AAV allow researchers...

...expressing Cas9 via transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells...Lines with Lentivirus Isolating a Monoclonal Cell Population by Limiting Dilution Lentivirus Production...performing and analyzing a gene edit using Addgene’s lentiviral CRISPR tools. Schematic overview of CRISPR...Cas9 expression in A549 cells transduced with lentiCas9-blast. Anti-Cas9 Western blotting was performed...of A549 cells transduced with Cas9 expressing lentivirus at the indicated multiplicity of infection (MOI...lines generated from A549 cells transduced with lentiCas9-blast. A549 cells were transduced (MOI = 37) and...PubMed PMID: 18007609. Addgene Materials Used 1. lentiCas9-Blast (Plasmid #52962) was a gift from Feng Zhang...

... the lentiviruses (like HIV-1) — that have been developed into lentiviral vectors. Lentiviruses are sometimes...Resources on addgene.org Addgene's Lentiviral Guide Most popular lentiviral plasmids γ-Retrovirus Guide References... 1: Gamma-retrovirus genome (left) and an HIV lentiviral genome (right.)  HIV seems an unlikely candidate...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 ...

...ready-to-use AAV and lentiviral preps! Download the green AAV background.       Some of our lentiviral plasmids ...Download the orange lentivirus background.         We also have ready-to-use lentiviral preps for COVID-...background.     If you’re working with AAV or lentivirus in your research, you might want to use one of...COVID-19 research, pooled CRISPR lentiirus libraries, pooled barcoding libraries, and more. Download the green...

...Mammalian Lentivirus pRDA_512 (RSV-driven; lentiGuide-based) Mammalian Lentivirus pRDA_722...722 (CMV-driven; lentiCRISPRv2-based; preferred over 512) Mammalian Lentivirus pRDA_789 (CMV-driven...Organism (Mammalian or drosophila) Delivery type (lentivirus, AAV, transposon, Plasmid) Destination vector...

...viruses grown in the lab are gamma retrovirus, lentivirus, adenovirus, and adeno-associated virus (AAV)...humans, earning this virus a BSL-2 classification. Lentivirus (a type of retrovirus) and gamma retrovirus require...and required in BSL-2 labs. When working with lentivirus, many organizations require or recommend disposable...you have to work with BSL-2 materials, such as lentivirus. Most of this work will need to be done in a ...that particles are not sprayed into the air. For lentivirus and other BSL-2 materials, you’ll need to use...for extra security. Finally, when spinning down lentivirus, you’ll need to use centrifuge bin covers, which...

...brain region of interest. AAV or lentivirus?  Both AAV and lentiviral vectors are used for intracerebral...wider diffusion is required. On the other hand, lentiviral vectors are superior packaging vectors for larger...astrocytes, and AAV4 targets mostly astrocytes. Lentiviral vectors are pseudotyped in most cases with VSV-G...injection. Find more details about choosing AAV or lentiviral vectors in our previous blog posts.  Viral vector...1013 vg/ml (vector genomes per milliliter) and lentiviral vectors around 108 IU/ml (infectious units per...

...Core, as well as pLV-dCas9-p300-P2A-PuroR for lentiviral expression. Figure 1: dCas9-p300 adds ...decrease methylation and activate transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD...Jaenisch’s lab in plasmid form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines...Tet-dependent (TetO-dCas9-D3A) constructs. For lentiviral expression, Fuw-dCas9-Dnmt3a and Fuw-dCas9-Dnmt3a-P2A-tagBFP...with the former also available as ready-to-use lentivirus. Figure 3: dCas9-DNMT3A methylates cytosines...hUbC-dCas9-MQ1(Q147L)-EGFP is also available for lentiviral transduction. Lysine-specific Demethylase 1 (...

... 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 ...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...

...digestion and ligation steps to build barcoded lentiviral plasmids containing one or more gRNAs. As each..., the ligation products are a diverse pool of lentiviral plasmids. Multiple rounds of digestion and ligation...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...

...individually label millions of mammalian cells through lentiviral infection and to monitor their abundance and ...optimize protocols for in vivo screening with lentiviral pooled shRNA libraries. Later, Bhang says, “When... treatment. The library consists of a pool of lentiviral vectors with roughly 73 million semi-random, ...infection. When using the ClonTracer system, lentiviruses containing these barcodes infect the target ... Find the ClonTracer Library at Addgene Find Lentiviral Plasmids at Addgene Check out Our Cancer Resources...

...efficiency. Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector...plasmids provide the necessary components to make lentiviral particles. Stable, tunable expression of Cas9...Difficulty Very easy Very hard Viral delivery   Lentivirus Retrovirus AAV Cells Dividing or nondividing...: Traffic Light Reporter System: Can evaluate lentiviral component delivery as well as genome repair by...

...the authors packaged it into lentivirus and used the resulting lentiviral library to generate a population...Science. 2013. PubMed PMID: 23328393 Wu lab human lentiviral CRISPR library     CRISPR pooled libraries enable...applications. First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed...Library. This complex second-generation CRISPR lentiviral library targets nearly all human protein-coding...from the CDC and cloned into a third generation lentiviral transfer vector. Each gene has been FLAG-tagged...dCas9-VPR limits one’s ability to package it into lentivirus and later create stable cell lines expressing... system has a much larger cargo capacity than lentivirus and is capable of efficiently creating stable...

...Cas9 activators can be delivered to the CNS by lentivirus, which has a larger packaging capacity than AAV...by the Day Lab, who developed neuron-optimized lentiviral vectors for in vitro and in vivo CRISPRa (Savell...Savell et al., 2019). This dual lentivirus system pairs ubiquitous expression of sgRNA(s) and mCherry (... such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression of sgRNA (top...neuronal physiology. The Day Lab’s neuron-optimized lentiviral vectors can be found here. High-throughput screening...

... 11. 3(3):346-53. Jaenisch Lentivirus Human Single polycistronic lentiviral vector for the expression ...May . 27(5):1042-9. Townes Lentivirus Human Single polycistronic lentiviral vector for the expression ...1221-9. Cibelli Lentivirus Human Single polycistronic, doxycycline-inducible lentiviral vector for the ...2):151-9. Jaenisch Lentivirus Mouse Polycistronic, doxycycline-inducible lentiviral vectors for the expression.... 106(1):157-62. Jaenisch Lentivirus Mouse Doxycycline-inducible lentiviral expression of mouse Oct4, ...7239):766-70. Nagy Lentivirus Mouse Polycistronic, doxycycline-inducible lentiviral vectors for the expression...Delivery Method Species Description Article PI Lentivirus Human Expression of human Oct4, Sox2, Klf4, Myc...

... those are lentiviral constructs. In 2007 we continued to see requests grow for lentiviral packaging and... pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with the pLKO ...-puro for retroviral expression and pLVTHM, a lentiviral plasmid for mammalian gene silencing. TALENs ...

...Base editors CRISPR toolkit for epitope tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery...toolkit   Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle Transient expression... Cas9. Recently, Baisong Lu’s lab developed a lentiviral capsid-based bionanoparticle system for packaging... aptamer binding protein for packaging into a lentivirus-like particle. Read the paper in Nucleic Acids...

... 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...collection of plasmids comprising the multiple lentiviral expression (MuLE) system from the laboratory ...the process of creating complex polycistronic lentiviruses. The MuLE toolbox contains a series of building-block... combinations of genetic elements in a single lentiviral vector provides a powerful tool to modify mammalian...routes in various conditions. The plasmids and lentiviral vectors encoding these proteins can also be used...

...different types of tubes: the ones we use to store lentivirus and the ones we use to store AAV. Our results...effect on the pH of the samples stored in our lentivirus tubes (Fig. 1). However, once thawed, there was... 1st Time CRISPR User) Tips for Titering Your Lentivirus Preps Additional Resources on Addgene.org Check... lentivirus...

...Blog Learn More about Lentivirus Figure out How to Get the Most from Your Lentiviral Transduction Additional... individually overexpressed and available for lentiviral packaging and delivery to target cells as a pooled... Fig. 1: Barcoded TF library was packaged in lentiviral particles and used to transduce human embryonic...

... A novel RNA targeting system Heat-inducible lentiviral Car constructs Anti-GFP [N86/8R] ready-to-use...of print. PMID: 36690762.   Heat-inducible lentiviral CAR constructs by: Brian O'Neill  A clever system...crux of the system is a set of heat-inducible lentiviral CAR constructs, the most basic of which is depicted..., inducible expression.   Find heat-inducible lentiviral plasmids here!   Wu Y, et. al. Control of the...

... CRISPR pooled libraries have been created in lentiviral systems for mammalian screens, which enables ...However, some cell types are poorly transduced by lentivirus, and thus screening with available CRISPR libraries...Primary murine T cells cannot be transduced by lentivirus and have been difficult to manipulate genetically...for screening of cells that are refractory to lentiviral transduction, now enabling forward genetic screens...

...conducted earlier by the Zhang lab for GeCKO, a lentiviral CRISPR library enabling genome-scale, knockout...that you review the steps for how to use pooled lentiviral CRISPR libraries and keep the following in mind... sequencing technology. These libraries are lentiviral in nature, so please ensure that you are equipped...equipped and authorized to make and use lentivirus. Also, note that due to the size of the backbone, transformations...

...12238 pLOX-CW-CRE Cre CMV Lentiviral Trono 12265 pHR-CMV-nlsCRE Cre CMV Lentiviral Trono 12493 p259 pCMV-...coexpression CMV Lentiviral Sadelain 30205 Cre-IRES-PuroR Cre and PuroR coexpression EF-1 alpha Lentiviral Kotton...-cre mCherry and Cre Lentiviral Luikart 66691 FUG-T2A-cre GFP and Cre Lentiviral Luikart 66692 pRubiC-...-iCre copGFP and iCre CB Lentiviral Oka 72257 pCDH-CB-iCre iCre CB Lentiviral Oka 73350 pAdx-CMV-iCre-... falxiparum Spielmann 86641 pLenti-hSynapsin-Cre-WPRE Cre hSyn Lentiviral Wang 86794 pCL20c-MSCV-IRES-...Eisenhoffer 131786 pLenti-CreLite CreLite system components, PhyBdeltaCreC and PIF6CreN, in lentiviral vector Cbh...Cre, Puro resistance and miRNA expression PGK Lentiviral Jacks 17736 pOG231 NLS-Cre CMV Mammalian Wahl...

...published a protocol and reagents for pseudotyping lentivirus with the SARS-CoV-2 spike protein. They deposited...19 pandemic, the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading frames...all but one are cloned into pLVX vectors (for lentiviral packaging) and contain an IRES-Puro marker for...(2020) Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization...

...parentheses.  In this example I wanted to find a lentiviral plasmid to express ubiquitin under control of...300+ plasmids. Under ‘Vector Types’ I selected ‘Lentiviral’ which eliminates most of the plasmids designed...this filter will allow to refine by serotype or lentiviral prep Requests: Only see plasmids with more than... cell type Serotype: Choose the AAV capsid or lentiviral prep needed Tags: Identify a specific marker ...

...AAV vectors collection. AAV Data Hub Lentivirus Production Lentiviral preparations are produced in 293T ...performed on lentiviral preparations that have been stored at -80 °C and thawed. Lentivirus titration is...for in our reported titers. Lentiviral vectors are titered using a lentiviral droplet digital PCR (ddPCR...viral production methods and quality control for lentivirus and AAV. Viral...AAV Production Titer Quality Control Resources Lentivirus Production Titer Quality Control Resources Overview... Cell culture medium (OptiPro SFM) containing lentivirus is first cleared by low-speed centrifugation ...cells are transduced with serial dilutions of a lentiviral vector, incubated for 72 h, and genomic DNA is...

...purpose. For mammalian lentiviral expression, we could use a vector like pLenti CMV Puro DEST (w118-1)...Gateway cloning to generate our desired constructs: lentiviral expression of the human KRAS gene in mammalian...available at Addgene, including the Frew Lab Multiple Lentiviral Expression Systems (MuLE) Kit, the MultiSite ...

...their options for tools such as empty backbones, lentiviral packaging systems, and luciferase expression.... Top Five by Views Guides CRISPR Guide Lentiviral Guide AAV Guide Optogenetics Guide γ-Retrovirus...a Liquid Bacteria Culture Collections Lentiviral Plasmids Tetracyline Inducible Plasmids Empty...help users get started with tools like CRISPR, lentivirus, and optogenetics — and shared the protocols ...

...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...

... NEB Stable   For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated...JM109-derived. For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated...Derived from HB101. For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated...cells for cloning and storage of unstable DNA and lentiviral/retroviral vectors. Used by Addgene for pooled...

...Description Element On or off PI 26429 pLenti CMV rtTA3 Blast (w756-1) Lentiviral reverse tetracycline-controlled...Resources: Browse Tetracycline Plasmids Tet-regulated Lentiviral RNAi Vectors Curated Plasmid Collections Background...On or Off PI 21916 Tet-pLKO-neo 3rd generation lentiviral plasmid for inducible expression of shRNA; neomycin... selection TetR On Wiederschain 41393 pCW57.1 Lentiviral vector for inducible expression for Gateway cloning...insert options rtTA On Root 11651 pLVUT-tTR-KRAB Lentiviral vector for inducible expression of transgene ...tTA or rtTA None On Vogelstein 25735 pSLIK-Neo Lentiviral vector for inducible expression of a miR-shRNA...third-generation rtTA On Fraser 44012 pInducer20 Lentiviral vector for shRNA expression; see article for ...

...., BMC Microbiology 2011 Feb 22;11:38. Lentiviral CRISPR libraries for knockout screening New systems...upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA delivery and providing...Pooled Libraries webpage or read our blog post,Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout...CoChR have been deposited to Addgene, including lentiviral and AAV expression vectors for certain variants...purification of Cas9 protein in Rosetta E. coli. New Lentiviral CRISPR activator and repressor plasmids from ...

...available in two vector formats: all-in-one (lentiCRISPRv2, expressing both Cas9 and an individual gRNA...gRNA from this single vector) and guide-only (lentiGuide expressing only an gRNA, for use with cells that...at Addgene, you can also order the individual lentiGuide gRNA plasmids targeting the human kinome. These...

...(Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem...systems which utilizes custom mouse strains and lentiviral expression vectors (Table 1). The four systems...pXPR_071, a plasmid that leads to higher titer lentivirus preparations than the first generation CHIME ...

...perform the transduction with: either retrovirus, lentivirus, adenovirus, or AAV. Then, your desired DNA to... some viruses integrate into the host genome (lentivirus and retrovirus) while others are non-integrating...Addgene.org Generating Stable Cell Lines with Lentivirus Virus Handling and Storage Addgene Viral Services...

...small but growing catalog of ready-made AAV and Lentiviral preps. This new Viral Service represents Addgene...establish consistent, high-quality protocols for lentiviral production. Bryan Roth’s lab from the University...the Addgene Blog Check Out the Top Requested Lentivirus and AAV of 2016 Learn How You Can Use the Viral...