...biosensor affinity for calcium.”  Primary Vector Type The vector type should describe how your plasmid ...indicate type in “Vector Type Other” column. For example, you may enter “donor template vector”. Cloning...difficult to grow. Growth Temp Choose from 30 °C, 37 °C, or room temperature. Growth Strain Please...plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival strain. For plasmids ...frog), D. rerio (zebrafish), D. melanogaster (fly), C. elegans (nematode), S. cerevisiae (budding yeast)... gentamicin, hygromycin, kanamycin, nourseothricin (clonNat), spectinomycin, streptomycin, tetracycline...maintain a glycerol stock. Choose from: ampicillin, apramycin, blasticidin, bleocin (zeocin), chloramphenicol...

...About Viral Vector Elements Learn How to Do Genome-Wide Screens with CRISPR Lentiviral Vectors Additional... by the envelop protein. Many times lentiviral vectors are engineered with non-native envelopes, a process... of oncoretroviral and lentiviral gene therapy vectors.” Biotechnology Advances 24.3 (2006). 321-37. Pubmed...titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009): 495-505. PubMed PMID...on the production of lentiviral and retroviral vectors.” Human Gene Therapy Clinical Development 25.3 ...Reiser, J. “Altering the tropism of lentiviral vectors through pseudotyping.” Current Gene Therapy 5.4...of the transductional efficiency of lentiviral vectors: effect of sera and polycations.” Molecular Biotechnology...

...modular vector assembly process designed for a quick, plug-and-play approach to cloning CRISPR vectors (McGee...for each module, generate the vector file, and quickly clone their vector of interest. And it truly is ...destination vectors for various delivery methods, along with compatible inserts. All destination vectors contain...heavy lifting out of CRISPR vector design, making it easier to get the vectors that are just right for your...Fragmid destination vectors currently available. Table 1: Fragmid destination vectors Organism Delivery...designing a CRISPR vector the Fragmid way!    Figure 1: Selecting your vector type on the Fragmid...plasmid and generating the vector file on the Fragmid website.   If the vector is to your liking, your...

...pCMVTNT PINK1 N-myc PINK1 Myc CMV Parkinson's Mark Cookson 13314 pCMVTNT PINK1 C-myc PINK1 Myc CMV Parkinson's...Ultra-Exon1Q23-Myc-A HTT Myc UbC Huntington's Baoji Xu 110487 Ultra-Exon1Q145-Myc-A HTT Myc UbC Huntington's...pUltra-Exon1Q23-Myc-B HTT Myc UbC Huntington's Baoji Xu 110489 pUltra-Exon1Q145-Myc-B HTT Myc UbC Huntington's...Strep-Lrrk2-Myc LRRK2 Myc, Strep CMV Parkinson's Maik Hintze 161583 pPuro3.1(+)_Strep-Lrrk2-Myc LRRK2 Myc, Strep...LRRK2-WT LRRK2 His, Myc CMV Parkinson's Ted Dawson 17612 pRK5-Myc-Parkin PRKN Myc CMV Parkinson's Ted ... pGW1-Myc-DJ1-WT PARK7 V5 CMV Parkinson's Mark Cookson 29349 pGW1-Myc-DJ1-L166P/K130R PARK7 Myc CMV Parkinson's...D1994S)-Myc LRRK2 Myc, Strep CMV Parkinson's Maik Hintze 161585 pPuro3.1(+)_Strep-Lrrk2(G2019S)-Myc LRRK2...

...Multi-luciferase reporter vector including transcriptional reporters for NF-kb, TGF-b, c-Myc, p53 and MAPK/JNK...regions Joshua Mendell 212936 pGL3 Basic Vector Firefly Vector for investigating regions controlling transcription...interest are cloned into a Firefly luciferase reporter vector via Gateway cloning to test for the presence of...pGL4.84(hRlucCP/Puro) RapidResponse(TM) Renilla Vector for investigating regions controlling transcription...Pete Stecha 212933 pGL4.84(hRlucCP/Puro) Renilla Vector for investigating regions controlling transcription...Mammalian expression of Nanoluc with a N-terminal Myc tag Erich Wanker 124701 pLenti-PGK-Venus-Akaluc (...expression of firefly luciferase Paul Khavari 109395 EFS-Vector Firefly EF1α AAV-CRISPR knockout plasmid. Firefly...

...Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This is the vector backbone. This...top requested plasmid from 2016 is an expression vector for CRISPR nuclease, SpCas9 from the Zhang Lab.... TLDR: This is a versatile mammalian expression vector that you can use to express both SpCas9 and your...Overall, this is a versatile mammalian expression vector that you can use to express both SpCas9 and your...mammalian cell lines. Alternative forms of this vector contain GFP-tagged and un-tagged versions of SpCas9...site). Finally, in addition to the 2A peptide on its C-terminus, SpCas9 is also fused to a 3X FLAG tag at...SpCas9 is fused to the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables...

...genome engineering methods for C. elegans were released, signaling a new era in C. elegans research. Homology... arms for your gene of interest into the FP-SEC vector using Gibson assembly. This construct will serve...of custom genomic modifications, is very robust in C. elegans, and the methods for HDR-mediated modification...the University of North Carolina has made CRISPR in C. elegans even easier - now, one can generate a fluorescent...-based gene tagging approach Multiple features of C. elegans have made CRISPR/Cas9 so successful in this...previously used a transgene selection strategy for their C. elegans/CRISPR experiments, but this strategy required... make it possible fluorescently label proteins in C. elegans, they inserted the SEC into a synthetic intron...

... A cis-splicing vector served as positive control (cis-ctrl). v1, vector 1; v2, vector 2; BD, binding ...SARS-CoV-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting microglia ...incorporated in an AAV vector (Figure 5). Efficiency and specificity of these AAV vectors for microglia was...human cells. Addgene offers vectors for CROPseq-multi using selection by Puromycin, Zeocin®, or nuclear expression...microglia The REVeRT Dual AAV Vector System Neurodegeneration Collection updates   Multiplexed perturbation ...Voltron2 voltage sensors By Brian O’Neill New viral vector preps expressing the Voltron2 voltage indicator...and Voltron2 are available as Cre-dependent AAV1 vectors and are suitable for extended imaging of neurons...

...El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, H., Kamuzinzi, R., Ledent...Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read our Blog... is to use a number of different bait and prey vectors. This has been shown to be as effective as using...organisms, including bacteria, alternative fungi (C. albicans, pC2HB) (7) and mammalian cells. If the ... prey libraries can be screened using both N- and C-terminal fusions of these proteins. This way both ...Two hybrid systems were developed in Saccharomyces cerevisiae in 1989 and are still used extensively ...

...into distinct expression vectors using the enzyme BbsI. These expression vectors each contain Type IIS restriction...Cas9 or dCas9-containing destination vectors. Destination vector options include humanized wt Cas9, dCas9...create lentiviral vectors expressing wt humanized Cas9 and up to three gRNAs. Entry vectors containing the...wt hCas9 entry vectors are supplied with the kit, you can clone your own entry vectors containing other...containing vector using Golden Gate assembly, and up to 8 gRNAs may be expressed simultaneously. Vectors for ...arrays (e.g. for spacers A, B, and C, you could obtain arrays A, B, C, AB, AC, BC, and ABC). Nielsen Lab...assembly of multiple components into a destination vector. The first step in CRISPR/Cas9 Golden Gate multiplexing...

...an ortholog in C. elegans making it ideal to study basic molecular biology processes. C. elegans are also...PMCID: PMC5525645. Mello, Craig C., et al. "Efficient gene transfer in C. elegans: extrachromosomal maintenance... al., 2009). By injecting mice with retroviral vectors that harbored different mutations found in leukemia...Gerald Rubin’s lab who created a set of modular vectors that can be used to optimally control gene expression...lab to study neuronal development (Brenner, 1973). C. elegans are small, transparent worms that have a ...life cycle and large brood sizes. The majority of C. elegans are also self-fertile hermaphrodites making... Nobel Prize in 2006. One of  major advantages of C. elegans is that they are transparent throughout their...

...visualized.  The tool-kit consists of highly useful vectors compatible with Gibson and Saptrap cloning systems...for delivering CRISPRi components by a single AAV vector are now available from Tonia Rex's lab.  vfCRISPR... Gether lab, now available as ready-to-use AAV vectors.   ...interactions New auxin inducible degron system for C. elegans New CRISPR plasmids New items from the viral...tool-kit for the auxin inducible degron (AID) system in C. elegans By Alyssa Cecchetelli The auxin inducible... degron (AID) system has been extensively used in C. elegans since 2015 to conditionally degrade proteins...repression, and editing with a single Cas9-VPR in Saccharomyces cerevisiae. To control Cas9 activity and influence...

... pREP4*, ** Kanamycin (pREP4) Cis-repression of the E. coli T5 promoter (found on vectors such as pQE ...lysozyme to reduce basal expression levels; expression vector cannot have p15A origin of replication F- ompT...higher T7 lysozyme expression than pLysS; expression vector cannot have p15A origin of replication F- ompT...codons not normally used in E. coli. Expression vector cannot have p15A origin of replication F- ompT ...alleviates inclusion body formation. Expression vector cannot have p15A origin of replication fhuA2 ...lac repressor on the pREP4 plasmid). Expression vector cannot have p15A origin of replication F-, Φ80ΔlacM15...conditions. Many recombinant proteins express better at 30°C or room-temperature, which is accomplished by growing...

... existing transposon and vector systems and creating an all-synthetic vector that included only the elements... into a destination vector in a predefined order. Finally, the destination vectors are used with Multisite... eight entry vectors can be used for each Multisite Gateway compatible destination vector for a maximum...step, compatible Level 0 vectors are directionally assembled into a Level 1 vector creating a single transcriptional...of interest into the pCoofy vectors, select the appropriate pair of vector and gene primers from Table...homology between the vector and gene primer for SLIC cloning. Amplify the pCoofy vector and the sequence ...recombination in SLIC cloning to form the desired vector. The pCoofy vectors contain the ccdB cassette, which is not...

...posts related to viral vector protocols and tips Read all of our viral vectors blog posts Resources on...calculate the viral titer: GC/mL = {[(R*C)(1000/V)]*D} R = Reaction volume C = Copies/uL V = Volume of virus ...Since Addgene began producing AAV vector preparations, we have used quantitative PCR to evaluate the ...evaluation of recombinant adeno-associated viral vectors." Human Gene Therapy ja (2019). PubMed PMID: 31140327...single-stranded and self-complementary adeno-associated viral vector genome titers by droplet digital PCR." Human gene...titration by qPCR Browse our collection of viral vector related protocols Learn more about the Addgene ...Song, Neng, et al. "Detection of circulating Mycobacterium tuberculosis-specific DNA by droplet digital...

...Cloning Vectors for Gram-Negative Bacteria. In R. L. Rodriguez & D. T. Denhardt (Eds.), Vectors (pp. 287... inform us on how to create better recombinant vectors. In this blog post, we’ll define plasmid host range...features to create broad host range recombinant vectors.  A plasmid’s host range is determined by the ...groups (Jain and Srivastava 2013).  Recombinant vectors that are made from broad host range plasmids are...   Building broad host range recombinant vectors Replicons, which consist of a plasmid’s ori and...range plasmids can be used to create recombinant vectors that can replicate in different bacteria and express...incorporating broad host range replicons, recombinant vectors can be modified further to increase the range of...

...yeast. These tagging vectors are based on previously described pFA6a-link vectors and include a Kan, SpHIS5...from these yeast-optimized fluorophore tagging vectors for your single or multi-color imaging experiments...substitution), and tagging at either the N- or C-terminus (3xHA, 13xMyc, GST, or GFP). Longtine et al.(3) describe...

.... (2007) ‘Improved Gateway Binary Vectors: High-Performance Vectors for Creation of Fusion Constructs ...are binary gateway vectors and can be used for promoter swapping and generating N and C terminally tagged...expression system Choosing an appropriate expression vector is the first step in generating transgenic Arabidopsis... ‘A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology’, Frontiers in Plant... plant selection markers (basta, hygromycin, kananamycin, tunicamycin) (Nakagawa et al., 2007). Alternatively...

...and improved. Takara’s “2nd generation Tet-Off vectors (Tet-Advanced)” are the most recent variants of...Recent work has even used all 3 in a single modular vector for ease of use! Conclusion This post contains ... Drosophila Terminators and PolyA Signals FLEx Vectors Resources at Addgene.org Tetracycline Collection...also shown that the Q system is also functional in C. elegans. Subsequent work by the Luo and Potter labs...23995928; PMCID: PMC3811519.Seo SO, Schmidt-Dannert C. Development of a synthetic cumate-inducible gene ...; PMCID: PMC5270762. Wendler, F., Park, S., Hill, C. et al. A LexAop > UAS > QUAS trimeric plasmid to ...genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197...

...Cepko 13779 pRho-Cre Cre-Myc rhodopsin Mammalian Cepko 13780 pNrl-Cre Cre-Myc, Expressed in rod photorecetor...Cre-ERT2 - Tamoxifen inducible; Gateway entry vector none Entry Vector Zon 27493 pBT140 Cre CMV Mammalian Luo...Cre-ERT2;Targeting vector for Fos locus Mammalian Luo 46389 pArcCreER Cre-ERT2;Targeting vector for Arc locus...Burgess 61570 Pvalb-2A-Cre targeting vector Cre; Targeting vector for Pvalb locus Mammalian Zeng 61573...61573 Rasgrf2-2A-dCre targeting vector DHFR-destabilized Cre; targeting vector for Rasgrf2 Mammalian Zeng ...Zeng 61574 Slc17a7-IRES2-Cre targeting vector Cre; Targeting vector for Slc17a7 locus Mammalian Zeng 62219...Nxph4-2A-CreERT2 Targeting Vector Cre-ERT2 - Tamoxifen inducible; Targeting vector Mouse Nxph4 Mammalian Zeng...