... for a researcher to come into contact with than GFP. Similarly, if the viral vector carries an shRNA ...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...

... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, ... prep # 55632-AAVrg). pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] was a gift from Edward Boyden (Addgene viral...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...

...selectable marker. Fluorescent reporters, such as GFP, are another common selection marker. Instead of ... sorting (FACS) is used to sort cells expressing GFP and later expanded into a cell line. More information... Guides Lentiviral Vector Guide Lentiviral Vector Guide Jump To: Lentiviral Vectors Lentiviral Generations...Read our lentiviral guide to learn about lentiviral components, generations, lentiviral production, and...Browse lentiviral plasmids available at Addgene. Lentivirus Versus Gamma-Retrovirus Lentiviruses and gamma-retroviruses... Things Lentivirus Lentiviral Vector Uses and Overview Overcoming the Challenges of Lentiviral Production...community for decades. Wild-type lentiviruses have been engineered into lentiviral vectors that can be safely...

...the viral protease, Nsp5. Our set also includes a GFP construct as a control. For easier portability, all...vector background, so you can package them into lentiviral vectors if you need to. Additionally, they have...

...Addgene has several great options: pLV hUbC-dCas9-T2A-GFP for removable dCas9 expression and pLM-CMV-R-Cre....One good system to balance these needs is the lentiviral-based expression of floxed dCas9 so that it may... stable gRNA expression can be achieved using lentiviral backbones expressing blasticidin or puromycin...

...cells in vivo, the non-immunogenic violet-excited GFP variant Vex. To understand the impact of this gene...high-throughput manner. They designed the pXPR_053 lentiviral expression vector, which contains an sgRNA insert...

...activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator helper complex with 2A GFP 61424 : sgRNA...(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). This is a AAV plasmid for sgRNA cloning. GFP-KASH fusion ...PX552; U6 promoter-driven; for sgRNA cloning; has GFP-KASH for FACS sorting SaCas9 + single guide RNA: ...system - lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9-Blast and lentiGuide-Puro (...#61425 - #61427 are intended for lentivirus production : 61425 : lentiviral vector encoding dCAS9-VP64 with... recombinase-2A-EGFP-KASH 60231 : sgRNA cloning backbone with Cre recombinase-2A-EGFP-KASH Detailed backbone...recombinase-2A-EGFP-KASH and an sgRNA. #60231 - AAV:ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR...

..., Hess et al. successfully evolved wild type GFP to EGFP using CRISPR-X and subsequent FACS sorting. They...with unique tissue-specific promoters expressing GFP, tetracycline response elements, and shRNAs many ... of DsRed connected to a pH-sensitive variant of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The...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...

..., Myc and SV40 Nuclear Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral...facilitates endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in...

...check out our article: Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection. Cellular...theoretically possible, but likely more challenging, with lentiviral expression. For more detail on how to design ...

...Vectors Read about Reporter Gene like Luciferase and GFP Resources on Addgene.org Find Plasmids for Your... as RNAP II promoters is commonly employed in lentiviral and retroviral constructs and we will discuss...

...activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can produce HIV-1 ...people worldwide are currently infected with the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral... with HIV-1. For two HIV-1 strains, Cas9/gRNA lentiviral expression significantly reduced the HIV-1 copy...PubMed Central PMCID: PMC4778041. Find CRISPR and lentiviral packaging plasmids used in this publication at...

... includes things like viral vectors that deliver GFP.   Figure 3: A) Cumulative AAV preps available...2016). Practical Considerations for Using Pooled Lentiviral CRISPR Libraries. Current Protocols in Molecular...

...vector Retroviral vectors: neomycin (pSIR-neo) , GFP (pSIR-GFP) , DsRed (pSIR-DsRed-Express2) , human CD2 (...amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9-Blast and lentiGuide-Puro packaging plasmids...sgRNA cloning lentiCRISPR v2 packaging plasmids: pVSVg , psPAX2 ; positive control: CMV-EGFP PDF, 2.4 MB...CRISPOR batch (Link opens in a new window) for lentiviral screens. sgRNA Scorer 2.0 (Link opens in a new...Musunuru CRISPRs in human pluripotent stem cells pCas9_GFP ; gRNA empty vector Protocol at StemBook O'Connor-Giles...plasmids: pVSVg , psPAX2 positive control: CMV-EGFP Pooled libraries are also available for human and mice...

...typically used for gRNA May contain reporter gene (e.g. GFP) to identify and enrich positive cells, or selection...transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich positive...components to make lentiviral particles (for details about lentivirus, see our Lentivirus Guide ) Stable,...high efficiency Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector...

...Calcium erGAP3 (GFP-Aequorin Protein) for imaging of Ca+ dynamics in endoplasmic reticulum GFP-Aequorin Protein...FlincG3 (GFP-based cGMP sensor) for imaging in C. elegans neurons Using a Robust and Sensitive GFP-Based ...2020 GENIE Project Calcium jGCaMP7 High-performance GFP-based calcium indicators (Constitutive or Cre-dependent...cyclic AMP) Signaling reporter island (SiRI) with GFP-based fluorescent reporter cAMPr Spatial Multiplexing... FLAMP Plasmids Jun Chu cGMP (cyclic GMP) FlincG GFP-based cGMP sensor Differential patterning of cGMP... vascular smooth muscle cells revealed by single GFP-linked biosensors. Proc Natl Acad Sci U S A. 2008...autophagic flux by pH-sensitive fluorescence (pMRX-IP-GFP-LC3-RFP) An Autophagic Flux Probe that Releases an...

...cassette comprised of OCT4, SOX2, LIN28, NANOG, and a GFP reporter. Minicircles are removed from cells by dilution...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...

...Cre-dependent expression of diphtheria toxin receptor (DTR)–GFP fusion protein. Azim et al. Nature. 2014 Apr 17. ... Proc Natl Acad Sci U S A. 2023 Aug 22. Use a lentiviral FRET-based reporter to study tau seeding in mammalian...

...typically used for gRNA May contain reporter gene (e.g. GFP) to identify and enrich positive cells, or selection...transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich positive...components to make lentiviral particles (for details about lentivirus, see our Lentivirus Guide ) Stable,...high efficiency Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector...

...selectable marker. Fluorescent reporters, such as GFP, are another common selection marker. Instead of ... sorting (FACS) is used to sort cells expressing GFP and later, sorted cells are expanded into a cell ...plasmids, as lentiviral vectors do. For more about lentiviruses, see Addgene's lentiviral vector guide...available at Addgene. Gamma-Retrovirus Versus Lentivirus Lentiviruses and gamma-retroviruses fall under the Retroviridae...main difference between lentiviruses and gamma-retroviruses is that lentiviruses are capable of infecting...that lentiviruses can infect a greater variety of cell types than retroviruses. Both lentiviruses and ...containing only the necessary packaging genes, while lentiviruses also contain accessory genes specific to each...