... methods is the expression of the reprogramming factors- OCT3/4, SOX2, KLF4, and C-MYC [1]. For the derivation...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 ... an episomal vector system that increases reprogramming efficiency through the expression of a shRNA against...RNA virus Episomal vectors- extrachromosomally maintained DNA plasmids that express reprogramming factors...Reprogramming efficiency of PBMCs with episomal vectors Episomal vectors are based on the Epstein-Barr Nuclear ...

...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 ...Addgene’s western blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the Applications...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...

...lentiviral expression of OSKM reprogramming factors: Mouse Expression Vectors Human Expression Vectors...harmful viral particles that express oncogenes, such as Myc. Viral vectors also have the largest genomic...Control Transgene Expression Mediated by a Sendai Virus-Based Vector Using a Nonstructural C Protein and Endogenous...Robert C. Robbins, Mark A. Kay, Michael T. Longaker, and Joseph C. Wu. "A nonviral minicircle vector for ... residual expression piggyBac Excisible Medium Medium Generates transgene-free and vector-free cells...so either a single polycistronic vector or multiple single-gene vectors can be used to package reprogramming...reprogramming. Additionally, lingering expression or re-expression of viral transgenes in iPSC-derived cells...

...of robust expression of high G+C content genes, as validated through monitoring the expression of fluorescent...different vectors and flow cytometry is used to sort populations by mCherry and GFP expression to check...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... be seen through the success of the change in expression of GFP, for the stop codon should have changed... can be applied for identification of essential MYC binding sites in a wide range of human cells.  A new...’s lab previously developed such a system in Streptomyces, a prokaryotic genus capable of producing a ...

...targeting vector (Fig. 1, Step 4), and co-injection of the targeting vector and a Cas9 expression plasmid...cell line stably expressing Cas9. This PCR toolkit offers C- and N-terminal tagging vectors (eg GFP, Flag...reaction to generate high-throughput targeting vectors. These vectors can be used to tag endogenous genes and...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)...mixed and transfected along with a Cas9 expression vector. They can also be introduced directly into a Drosophila...PCR or cloning, as digestion of the destination vector with SapI yields 2 sites- the first site accepts...

...cerevisiae expression vectors are available at Addgene. A set of mammalian expression vectors containing... be easily expressed in non-mammalian cells. Yeast expression plasmids The yeast expression constructs....  Bacterial expression plasmids For those constructs to be used in bacterial expression systems, Ginkgo...backbones which drive expression using either TEF1 or GAL1 promoters. These vectors are intended for the...tag, cleavage, codon usage, and expression system. Mammalian expression plasmids Finally a set of wild ... (spike protein) cloned into a CMV driven expression vector are also available. These contain no tags ...coding genes in E. coli and S. cerevisiae expression vectors in late March. The first set of 383 constructs...

...stage involves overexpression of four reprogramming factors- OCT4, SOX2, KLF4, and C-MYC - that induce ...additional pluripotency stage) Less Safety Less (C-MYC, a known oncogene is one of the reprogramming factors...four reprogramming factors can be delivered and expressed in multiple somatic cells via various methods....transdifferentiated in vivo to pancreatic β-like-cells with the expression of pancreatic and duodenal homoeobox gene1 (PDX1...transdifferentiation is usually carried out by expressing transcription factors specific to the lineage...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...

...Fluorescent Protein (GFP) Fluorescent Protein Guide Expression Vectors from the Berkeley QB3 MacroLab Eric thanks...to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses of the affinity tag...box of tags and fusion proteins to aid in the expression and purification of recombinant proteins. Tags...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... tags, a His tag can be fused to either the N- or C-terminus of a protein. Unlike other epitope tags –...Biochemists and molecular biologists who need to overexpress and purify proteins can face any number of technical...

...using FX cloning) into the expression vector pSb-Init, which attaches a Myc-tag and a His6-tag to the ...protocol starts out with anti-Myc antibodies bound to an ELISA plate to capture Myc-tagged sybodies. Then, the...their small size, nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target...sybodies can be subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for the production of tag-free proteins...due to camelid high body temperature (37.2 - 39.2°C). The Seeger lab found a way around this by using ...

...cells) through the expression of transcription factors OCT3/4, SOX2, KLF4, and c-MYC is the established...low off-target activity; mammalian expression vector Rees H, et al. Nature Communications. 2017. PubMed...and ribozyme systems for multiplex gRNA expression. Expression of toolkit components can be controlled...gene editing. The bacterial and mammalian expression vectors for these CRISPR inhibitors are listed in...Species Bacterial Expression Construct SpCas9 Inhibition (E. coli) Mammalian Expression Construct SpCas9...- and C-terminal tags for mTagBFP, TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and empty vectors that have...neuronal tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community...

... single protein is expressed, with the intent of purification, it will be expressed along with all other...not naturally exist in most expression systems that can be added to the N or C terminus of your protein ...other cellular proteins within the expression system (bacteria, yeast, mammalian, etc.). To individually...about that protein that no other protein in the expression system has. This is where affinity tags come ...on: The protein you are trying to purify The expression system If you want the tag to remain on your ...MBP tag, although large, can actually increase expression and solubility of the tagged protein (Fox and...different types of tags, including epitope tags (FLAG, Myc, etc.) and fluorescent protein tags (GFP, mCherry...

...bacterial expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT...2xNLS) and c-terminal (Cas9(C)-2xNLS) fragments facilitates target DNA cleavage upon co-expression and spontaneous...destination vector expressing a separate reporter gene (EGFP, iRFP, IFP1.4, puromycin, neomycin or luciferase...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ... SFFV) or inducible (CMV-TO) cDNA expression, sgRNA (U6) expression, shRNA/shRNA-miR30 constitutive (H1...-miR30 (CMV-TO) expression, or a promoterless construct. Pre-constructed Entry vectors containing Cas9...a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed with the...

...escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in human cell lines...Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization Signals, and GFP. B...structured illumination microscopy in HeLa cells expressing COX8a-mStayGold. Scale bars 10 μm (main), 1 μm...developed an engineered adeno-associated virus (AAV) vector named AAV.CAP-Mac for non-invasive systemic gene...bias in mature marmosets. CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured...cultured human neurons, with broader neuronal expression than AAV9. This new capsid will be a valuable ...N.C., Goeden, N. et al. Adeno-associated viral vectors for functional intravenous gene transfer throughout...

... 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...protein expression constructs fused with epitopes or fluorophores, generate expression vectors with various...AAV transfer vector for in-vivo expression and plasmids optimized for dCas9-VPR expression in drosophila...parasites that constitutively express Cas9 with a library of sgRNA expression vectors (containing ten guides ... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ... to test multiple expression systems before they achieve the appropriate expression level for their gene...

...epitope tags, low FP expression may be insufficient for monitoring a weakly expressed POI. Spaghetti Monster...cytotoxicity, low immunogenicity, and the long-term expression of AAV delivered genes. AAV serotypes have been.... Thanks to this work it is now much easier to express molecules in your favorite neuronal population ...hemagglutinin (HA), myelocytomatosis viral oncogene (myc), simian virus 5-derived epitope (V5), the synthetic...insufficient for detection when the POI is weakly expressed. As these epitopes tags have no intrinsic visual... be fused to scaffold proteins in order to be expressed stably in neurons. As compared to epitopes tags...labeling of neurons. As discussed above, weakly expressed proteins in particular are difficult to track ...

... 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...Transducibility Some viral vectors are tested in vitro and in vivo for gene expression and/or function. These...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 ...

...(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... (although we could successfully knock-in non-expressed genes with similar efficiency, Dewari et al. 2018...tagging success, although many of these genes were expressed at really low levels so actual knock-in success...available for human and mouse genomes only and deals with C-terminus epitope tagging. If you work with other model...

...transfected with both vectors. Flex vectors are the solution. Flex vectors expressing fluorescent cytoplasmic...concentrations of vectors expressing the required recombinase. The high concentration of FLEx vectors guarantees.... It became possible to design FLEx vectors (Flip-excision vectors) that only allow one inversion event...types in neuronal networks Double inverted vectors expressing light inducible ion channels (plasmid # 20298...cell specific manipulations in vivo, FLEx vectors expressing the light activatable ion channel under the...optimum of 30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular...specific) control of DNA recombination and gene expression can be advanced by the coordinated use of fellow...

...luciferase, cre/lox, yeast expression, worm expression, insect expression, plant expression, mouse targeting, ...biosensor affinity for calcium.”  Primary Vector Type The vector type should describe how your plasmid ...to be used. Choose from: mammalian expression, bacterial expression, lentiviral, retroviral, AAV, RNAi...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)...

...About Viral Vector Elements Learn How to Do Genome-Wide Screens with CRISPR Lentiviral Vectors Additional...becomes cloudy. Mycoplasma compete with host cells for nutrients and can alter expression of receptors, ..., Margison G.P. “Mycoplasma infection significantly alters microarray gene expression profiles.” Biotechniques...often employed to establish cell lines that stably express a gene of interest. While the general approach ...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting...on a variety of factors such as the transgene expressed, the viral storage buffers used, and other experimental... by the envelop protein. Many times lentiviral vectors are engineered with non-native envelopes, a process...