... Viral vectors are prone to recombination In lentiviral vectors, recombination often occurs between the...time, bacteria containing the recombined vector backbone will also outgrow bacteria that contain the full...full length plasmid.   Figure 1: The full lentiviral plasmid is expected to be >12 kb. However if ...smaller plasmids. The ~3.7 kb recombined vector backbone contains the antibiotic resistance gene (AmpR)...noticed that in Stbl3 bacterial cultures with lentiviral vectors, the flat white colonies contained the...E. coli cultures contain the recombined vector backbone. Test multiple colonies before saving a glycerol...different band patterns for the recombined vector backbone vs. the full length plasmid. If you find that ...

...3rd generation lentiviral vector backbone from the Shih lab. You can find more lentiviral expression vectors...descriptions of the lentiviral elements and the 2nd and 3rd Generation Lentiviral Systems can be found...found in our Lentiviral guide. Popular packaging and lentiviral envelope plasmids are listed here. Please...pre-treated with a compound of interest. Lentiviral vectors Popular Lentiviral Transfer Vectors    Expressing your...Plasmid #52961) This is a 3rd generation lentiviral backbone from the Zhang lab expressing S. pyogenes...in the genome editing field. Lentiviral Plasmid Kit Multiple Lentiviral Expression System Kit (Kit #1000000060...Murine Stem Cell Virus (MSCV) genomes whereas lentiviral vectors are derived from the human immunodeficiency...

... those are lentiviral constructs. In 2007 we continued to see requests grow for lentiviral packaging and... and empty backbones such as pBABE-puro for retroviral expression and pLVTHM, a lentiviral plasmid for...2006, we saw an increase in popularity of empty backbones for shRNA expression, such as pLKO.1 puro and ...and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with the pLKO plasmids you'll...

...Addgene's most popular lentiviral plasmids Overview of Lentiviral Packaging Plasmids Lentiviral Protocols and ...frequently asked lentiviral questions. This useful resource from Addgene Depositor (and lentiviral expert) Didier...plasmid backbone that was designed specifically for transient expression (find some useful backbones here...HIV-derived. Get Tips on Titering Your Lentivirus Concerning safety of lentiviral transduction Q8: You mentioned....  Addgene has an extensive collection of lentiviral plasmids created for a variety of applications including... Didier Trono's lab webpage explains how lentiviral particles are produced by co-transfecting three essential...essential ingredients: the lentiviral packaging vector, a transfer vector including the gene of interest...

...sequence. Empty backbone A plasmid designed for cloning a gene/insert into it. Empty backbones often contain...present in the backbone or was a part of the insert (when cloned into the vector backbone). If your plasmid...appropriate boxes with the vector backbone manufacturer, vector backbone size, total size of your vector...made to the backbone, and the vector type. Pro Tip! Reference the Addgene ID in the "Backbone Manufacturer...categories (e.g., for a lentiviral vector to be used in mammalian cells, check Lentiviral and Mammalian Expression...library is a set of plasmids all built with the same backbone and only differing in the gene/insert. Pooled ...Insert *Note: This page will not appear for empty backbones Please enter the name of any gene contained within...

... vector backbones; once validated these universal primers can be used to titer any lentiviral prep that...considerations for titering lentivirus Volume and transduction time will influence lentiviral titers as will the...defective viral particles. Direct measurement of lentiviral RNA is one alternative to direct p24 measurement... for functional assays depending on the vector backbone. When using qPCR based methods for viral titration...transgene expression. The most accurate method of lentiviral titration uses qPCR to measure the number of ...; the titer measured with a freshly collected lentiviral prep may be significantly higher than the same...concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009): 495-505...

... Plasmid Collections Empty Backbones Choosing Your Perfect Plasmid Backbone Background When choosing what... Backbones Neomycin (G418) Mammalian Tet-pLKO-neo - Tet-inducible lentiviral shRNA...vector - Lentiviral shRNA expression Tet-pLKO-puro - Tet-inducible lentiviral shRNA expression...Tet-inducible lentiviral shRNA expression pLenti CMV Hygro DEST - Lentiviral Gateway destination...expression through lentiviral delivery pLenti CMV GFP DEST - Lentiviral Gateway destination...Advantages Representative Empty Backbones Lentiviral Can transduce both dividing and nondividing...A guide to Addgene's empty vector backbones curated by species, epitope tags/fusion protein, selectable...

...designed to have increased lentiviral titer as compared to previous lentiviral vectors. It can be used both... the Addgene Website Visit our Lentiviral Guide Pages Find Lentiviral Plasmids Find CRISPR Pooled Libraries... lentiCRISPR v2 as the backbone for their many gRNAs. The high titer lentivirus production afforded by...purpose As its name suggests, lentiCRISPR v2 is a lentiviral transfer vector used to deliver both Cas9 and...

...Lab pCMV-VSV-G (Empty Backbone) Envelope protein for producing lentiviral and MuLV retroviral particles...with a packaging vector such as pCMV-dR8.2 dvpr (lentiviral) or pUMVC (MuLV retroviral). Bob Weinberg ...

...mammalian expression system and go for a lentiviral or a retroviral backbone. This is considered to be a more ... empty backbones for additional details and guidance for choosing your perfect plasmid backbone. Or just...Vectors (or empty backbones) are frequently used in molecular biology to isolate, multiply, or express...At Addgene, we have a vast collection of empty backbones that have been designed, tested, and published... empty backbone...

...Calabrese in backbone XPR_502 (P65 HSF), Set A Human CRISPRa sgRNA activation library in backbone XPR_502 ...blasticidin resistance from EFS promoter. Lentiviral backbone. Zhang Cas9 and Accessories for Activating...available in lentiviral format. Viral...Brunello in lentiGuide-Puro Human sgRNA library in backbone lentiGuide-Puro targeting 19,114 genes and containing...sgRNAs along with 1000 non-targeting controls. This backbone does not contain SpCas9. 76,441 unique sgRNAs ...library in lentiCRISPRv2 Human sgRNA library in backbone lentiCRISPRv2 targeting 19,114 genes and containing...sgRNAs along with 1000 non-targeting controls. This backbone contains SpCas9. SpCas9 and 76,441 unique sgRNAs...

...the plasmid backbone is meant to be descriptive of the components found within the backbone. For example... well as the viral LTRs, differ. As a result, lentiviral and γ-retroviral packaging plasmids are not interchangeable...using 293T cells This method is very similar to lentiviral packaging methods. The three components described...found within both the 5' and 3' LTRs of retro/lentiviral plasmids. U5; Unique 5'; region at the 5' end... cells a virus can infect. Note: While both lentiviruses and γ-retroviruses use the same gene products...How do engineered γ-retroviruses differ from lentiviruses? There are a few important differences between...between engineered γ-retroviruses and lentiviruses. First, these viruses were derived from different genomes...

...the EF-1a promoter. It’s a 3rd generation lentiviral backbone that must be used with lentiCas9-Blast or...the last 15 years - the majority of these being lentiviral or CRISPR tools. How many times have these plasmids...Addgene psPAX2  - psPAX2 is a 2nd generation lentiviral packaging plasmid from the Didier Trono lab that...genes. It can be used with 2nd or 3rd generation lentiviral vectors and an envelope expressing plasmid, such...PX458). lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used...and contains an envelope protein for producing lentiviral and MuLV retroviral particles. It can be used...with packaging plasmids such as pCMV-dR8.2 dvpr (lentiviral) or pUMVC (MuLV retroviral). Fun fact: published...

...Mammalian/Lentiviral BbsI none S. pyogenes Puro, TagBFP Yusa lentiGuide-Puro 52963 Mammalian/Lentiviral BsmBI... Puro Zhang lenti sgRNA(MS2)_zeo backbone 61427 Mammalian/Lentiviral BsmBI none S. pyogenes Zhang pL-CRISPR.EFS.GFP...Mammalian/Lentiviral BfuAI none S. pyogenes Puro Wolfe pL-CRISPR.EFS.tRFP 57819 Mammalian/Lentiviral BsmBI...Mammalian/Lentiviral BsmBI none S. pyogenes ccdB Wei pLKO5.sgRNA.EFS.PAC 57825 Mammalian/Lentiviral BsmBI ...Mammalian/Lentiviral BsmBI yes, cut S. pyogenes Puro Ebert pL-CRISPR.SFFV.PAC 57829 Mammalian/Lentiviral BsmBI...62348 Mammalian/Lentiviral BbsI none S. pyogenes Puro Ochiai pSB700 64046 Mammalian/Lentiviral none S. pyogenes...57822 Mammalian/Lentiviral BsmBI none S. pyogenes EGFP Ebert pSECC 60820 Mammalian/Lentiviral BsmBI yes, cut...

...important in disease. Empty Backbones This table is a collection of empty backbone plasmids into which you...with Nano-lanterns Collection Highlights Empty Backbones Expression Constructs Reporter Constructs Luciferase...regulatory elements on gene expression. Empty backbones for inserting regulatory elements before or after...promoter/enhancer regions using Gateway cloning. Lentiviral. Jorge Ferrer 87067 pcDNA3.1-ccdB-Nanoluc NanoLuc...N-terminal Nanoluc fusions using Gateway cloning. Lentiviral. Mikko Taipale 12563 PCD FL0X Firefly Insertion...Firefly luciferase fusions using Gateway cloning. Lentiviral. Mikko Taipale 87077 pLenti6.2-Firefly-ccdB Firefly...Firefly luciferase fusions using Gateway cloning. Lentiviral. Mikko Taipale 87789 pPTluc Firefly Insertion...

... for using pooled lentiviral libraries. A) Amplification, validation, and lentiviral generation of a pooled...Usage of a pooled library lentiviral prep for experiments. Pooled library lentiviral preps are generally aliquoted...Pooled Lentiviral CRISPR Libraries- A nice overview of what to consider when using pooled lentiviral CRISPR...production of lentivirus. Purification of this lentivirus results in the pooled library lentiviral prep. B).... Most of the pooled libraries at Addgene are lentiviral pooled libraries, though other types exist.  ...plasmids requires amplification, verification, and lentiviral production steps before you can start transducing... After functional titer is determined, larger lentiviral preps can be used for screening-scale transduction...

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

...CombiGEM-based assembly into the CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28...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... pAWp28 storage vector to the pAWp12 lentiviral vector should retain the diversity in your gRNA library...recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library." Nature biotechnology...

...Stabilizing gRNA modifications  Sugar phosphate backbone modifications The recognition sequence for CRISPR-Cas9...modify the sugar or phosphate molecules of the RNA backbone to increase stability.  Nature has evolved ways...replaces a nonbridging phosphate oxygen. Other useful backbone modifications to consider include amide linkages...and a constrained ethyl.  Can’t decide which RNA backbone modification is best? Studies suggest coupling...Request ready-to-use viral preps of select CRISPR lentiviral plasmids Additional resources on the Addgene...

...to work with. For example, a library in a lentiviral backbone might be more prone to recombination, while...consider if there are any features of the library backbone that would make the library more difficult to ...while a library with a large backbone might be more difficult to transform.    Figure 1: Factors...other colonies containing smaller, recombined backbones that may replicate faster. For this reason, if...