...generated a mixture of two vectors that encoded six factors - OCT3/4, SOX2, KLF4, C-MYC, LIN28 and SV40 large...enhances iPSC generation as does replacing C-MYC with L-MYC [10, 11]. While the mechanisms underlying ...addition of an shRNA for P53 and replacing C-MYC with L-MYC in the episomal reprogramming cocktail by ...Reprogramming efficiency of PBMCs with episomal vectors Episomal vectors are based on the Epstein-Barr Nuclear ... reprogramming factors- OCT3/4, SOX2, KLF4, and C-MYC [1]. For the derivation of clinical grade hiPSCs...purposes. In this post, I’ll describe an episomal vector system that increases reprogramming efficiency ...modification of the original genome by integrating viral vectors are preferred. This is to avoid the chances of ...

...on the Anti-c-Myc [9E10] page for more details. Image credit: Addgene.   Find anti-c-Myc antibody here...the C-terminal region of the human proto-oncogene c-Myc. Since the generation of the first anti-c-Myc ...light-gated potassium channelrhodopsins Myc-tag antibody anti-c-Myc [9E10] now available! Improved voltage...doi.org/10.1101/2021.09.17.460684.)   Myc-tag antibody anti-c-Myc [9E10] now available! by: Ashley Waldron...to guess that the Myc-tag is one of them (and not just because of the title!) The Myc-tag is a string of...), the Myc tag has become one of the most commonly used protein tags - a quick search for ‘Myc tag’ in... of Addgene’s western blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the...

...genome-wide. It was designed based on MYC-ChIP-seq data from several MYC-dependent cancer cell lines. It can...selection marker, Thy1.1, in addition to puromycin resistance.  The MYC-CRISPR library targets E-boxes genome-wide... a) Cells are transfected with three different vectors and flow cytometry is used to sort populations ... can be applied for identification of essential MYC binding sites in a wide range of human cells.  A new... system is capable of robust expression of high G+C content genes, as validated through monitoring the...’s lab previously developed such a system in Streptomyces, a prokaryotic genus capable of producing a ...

...Robert C. Robbins, Mark A. Kay, Michael T. Longaker, and Joseph C. Wu. "A nonviral minicircle vector for ...particles that express oncogenes, such as Myc. Viral vectors also have the largest genomic footprint of...so either a single polycistronic vector or multiple single-gene vectors can be used to package reprogramming...reprogramming vectors available from Addgene. Another drawback is that lentiviral vectors insert in different...Cre-lox lentiviral vectors: This approach is the same as using a standard lentiviral vector, but transient...Non-integrating iPSC delivery methods Viral Vectors Adenovirus: Adenoviral vectors infect dividing and nondividing...reprogramming factors: Mouse Expression Vectors Human Expression Vectors Carey et al Hockemeyer et al Brambrink...

...reaction to generate high-throughput targeting vectors. These vectors can be used to tag endogenous genes and...the final targeting vector (Fig. 1, Step 4), and co-injection of the targeting vector and a Cas9 expression...site destination vectors for inserting a tag at multiple target sites. SapTrap vectors can be easily modified...cells. To use CRISPaint, users need 3 vectors:  1) a gRNA vector that targets the gene of interest, 2)...expressing Cas9. This PCR toolkit offers C- and N-terminal tagging vectors (eg GFP, Flag, YFP, Strep, TEV-V5)...PCR or cloning, as digestion of the destination vector with SapI yields 2 sites- the first site accepts...specify the reading frame of the insertion, and 3) a vector containing the desired tag, which can be obtained...

...to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses of the affinity tag...step purification with amylose, but wicked huge Myc EQKLISEEDL  1.2 Affinity Frequently used for western...purification to get your protein as pure as possible? Vectors that offer different combinations of tags are readily...Protein (GFP) Fluorescent Protein Guide Expression Vectors from the Berkeley QB3 MacroLab Eric thanks his... tags, a His tag can be fused to either the N- or C-terminus of a protein. Unlike other epitope tags –...

...four reprogramming factors- OCT4, SOX2, KLF4, and C-MYC - that induce a differentiated somatic cell to revert...additional pluripotency stage) Less Safety Less (C-MYC, a known oncogene is one of the reprogramming factors...and in vitro methods are similar except that the vectors carrying the transdifferentiation factors are directly... (iPSCs) by temperature-sensitive Sendai virus vectors. Proc Natl Acad Sci U S A, 2011. 108(34): p. 14234...., Human induced pluripotent stem cells free of vector and transgene sequences. Science, 2009. 324(5928...mouse induced pluripotent stem cells without viral vectors. Science, 2008. 322(5903): p. 949-53. PubMed PMID...Differentiate the Stem Cell Field Learn how lentiviral vectors can be used for gene delivery Resources on Addgene.org...

...FX cloning) into the expression vector pSb-Init, which attaches a Myc-tag and a His6-tag to the end of...protocol starts out with anti-Myc antibodies bound to an ELISA plate to capture Myc-tagged sybodies. Then, the...due to camelid high body temperature (37.2 - 39.2°C). The Seeger lab found a way around this by using ...

...cerevisiae expression vectors are available at Addgene. A set of mammalian expression vectors containing various...sites (thrombin and TEV), and C-terminal tags (StrepII, 3xFLAG, 6xHIS, MBP, Myc). These constructs include...expression using either TEF1 or GAL1 promoters. These vectors are intended for the high level expression and ...spike protein) cloned into a CMV driven expression vector are also available. These contain no tags and are... genes in E. coli and S. cerevisiae expression vectors in late March. The first set of 383 constructs ...

...- and C-terminal tags for mTagBFP, TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and empty vectors that have...of transcription factors OCT3/4, SOX2, KLF4, and c-MYC is the established model for reprogramming somatic...neuronal tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community...discrepancies. The polycistronic piggyBac transposon vectors tested in this work are available at Addgene. ...for the research community. You can find the AAV vectors for infecting GABA-ergic interneurons here. Dimidschstein...Very low off-target activity; protein purification vector  87439  pCMV-HF-BE3  HF-Cas9 (NGG)  Very low ...low off-target activity; mammalian expression vector Rees H, et al. Nature Communications. 2017. PubMed...

...different types of tags, including epitope tags (FLAG, Myc, etc.) and fluorescent protein tags (GFP, mCherry...Ready to start tagging? We are here to help with vectors, protocols, antibodies, and more!   Resources ... expression systems that can be added to the N or C terminus of your protein of interest. Many of these...

...Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization Signals, and GFP. B...developed an engineered adeno-associated virus (AAV) vector named AAV.CAP-Mac for non-invasive systemic gene...N.C., Goeden, N. et al. Adeno-associated viral vectors for functional intravenous gene transfer throughout...region VIII of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial cell-specific tropism, with... by pre-existing antibodies against AAV9-based vectors (and vice versa), enabling repeat dosing. In a ...mChe), with co-incubation with assist peptide (AP). C) Workflow for Cas-RNP-PAGE editing of primary T cells...

...expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT vector ensures...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ...custom-cloned vectors in a Multisite Gateway reaction with a desired destination vector expressing a separate...MXS-chaining kit contains 5 empty chaining vectors plus a destination vector and a collection of building blocks...different N- and C-terminal tags. Additionally, by using an adapted destination vector with a heterologous... Cas9 into n-terminal (Cas9(N)-2xNLS) and c-terminal (Cas9(C)-2xNLS) fragments facilitates target DNA ...activation, the c-terminal cas9 fragment was fused with FK 506 binding protein 12 (Cas9(C)-FKBP-2xNLS) and...

...(3xFLAG, Myc, V5, or HA). Tag-specific PCR genotyping can tell if HDR completely failed. If C-terminus...Kalantzaki M, Gangoso E, Matjusaitis M, Garcia-Diaz C, Blin C, Grant V, Bulstrode H, Gogolok S et al. 2017. ...pitfalls: they require construction of plasmid vectors and/or rely on cumbersome in vitro synthesis of...purchased and there is no need to construct plasmid vectors or synthesize/purify IVT sgRNA in the lab. Figure...available for human and mouse genomes only and deals with C-terminus epitope tagging. If you work with other model...your crRNAs using other web-based design tools (for C-terminus tagging, pick a crRNA that cuts 8-15 bp downstream...insertion site (within ~8-15 bp from the stop codon for C-terminus tagging) GC content of the crRNA (aim for...

... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core. Transfections...produced the viral vector lot. Contact us to determine which facility produced your viral vector lot. At Addgene...Empty vector particles can be identified after negative staining and appear darker than full vector particles...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...-80 °C. Titer Titering is either performed by Addgene or by the University of Pennsylvania Vector Core...80 °C. Titer All titering is performed on lentiviral preparations that have been stored at -80 °C and ...method.  Titering by the University of Pennsylvania Vector Core is (as of April 2016) also performed by ddPCR...

... 79628 (C) Cv (capsid) N and C 79633 (N) and 79641 (C) NS1 (var: W98G) N and C 79635 NS3 C 79637...79637 NS2B C 79636 NS4A N 79640 NS4B C 79639 NS5 C 79632 (N) and 79631 (C) PrM N and C   Detecting...expression vectors with various promoters suitable for in vivo expression, and/or produce vectors to make...prototrophic strains. The vectors were based on the original EasyClone vectors (Jensen et al 2013) allow...donor vectors containing mAID and either Neo or Hygro resistance markers with CRISPR/Cas vectors targeting...tagged BioID2 for N-terminal fusions and Myc tagged BioID2 for C-terminal fusions. Kim, et al. Mol Biol... interest and cloned into provided destination vectors using low-cost, one-pot golden gate cloning that...

...hemagglutinin (HA), myelocytomatosis viral oncogene (myc), simian virus 5-derived epitope (V5), the synthetic...Spaghetti Monster probes at Addgene Browse all AAV vectors Find ready-to-use AAV preps ...pdb-top63. PubMed PMID: 20150100. 4. Shaner, Nathan C., George H. Patterson, and Michael W. Davidson. "Advances...

...are transfected with both vectors. Flex vectors are the solution. Flex vectors expressing fluorescent cytoplasmic.... It became possible to design FLEx vectors (Flip-excision vectors) that only allow one inversion event...concentrations of vectors expressing the required recombinase. The high concentration of FLEx vectors guarantees...optimum of 30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular...experiments. Mutated FRT and loxP sites in Flex Vectors – inversion only once All recombination events ...between target sites of the same sequence in FLEx vectors, the DNA sequence that should be inverted is flanked... target sites are cleaved out, leaving behind a vector with the inverted DNA flanked by non-compatible...

...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...