...difficult-to-manipulate primary cells, achieving knockdown within 1-2 hr of reagent delivery. TRIM-Away is suitable for... inducer levels. That is, if transcription factor 1 controls expression of transcription factor 2, which...modulating the induction of transcription factors 1 and 2. See Aranda-Díaz et al. for more information...interest. These transcription factors consist of the GAL4 or ZIF268 DNA binding domains fused to either the...fluorescent reporters and other proteins controlled by GAL4 and ZIF268. To demonstrate independent control of... Aranda-Díaz et al transformed yeast with both a GAL4/estradiol controlled transcription factor and a ...

...of an industry H-1B need to be aware of the April 1 deadline for companies to submit an application on...and non-profit organizations do not have the April 1 H-1B queue process or quota like industry, these organizations... National Interest Waiver. The three prongs are: (1) The foreign national’s proposed work endeavor must...-1B, you need to prove 3 threshold requirements: (1) that you have international recognition for your ...provide legal advice or opinions. The contents of this post should not be relied upon for legal advice ...information presented may not reflect the most current legal developments.  Further, this post may contain technical...attorney should be contacted for advice on specific legal issues.  The permanent residence “green card” process...

...tools in Drosophila (Fig. 1):  GAL4-UAS, a transcription factor binding site. GAL4 expressed in the fly binds....  Genetics of SPARC Figure 1: SPARC and SPARC2 use the GAL4-UAS system along with precisely truncated...

...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6...transducing cells: galactose and the AAV receptor (AAVR). Cell lines lacking galactose were still transduced...PHP.B, unlike its parental AAV9, does not require galactose and AAVR to transduce cells. The Wilson Lab generated...

...supE44 thi-1 gyrA96 relA1 tonA JM109 Addgene; Promega e14-(McrA-) recA1 endA1 gyrA96 thi-1 hsdR17(rK- ...England BioLabs E. coli B F dcm ompT hsdS(rB mB) gal ccdB Survival Invitrogen F- mcrA Delta(mrr-hsdRMS-mcrBC...mB–) recA13 supE44 ara-14 galK2 lacY1 proA2 rpsL20 (StrR ) xyl-5 λ– leu mtl-1 Top10 Invitrogen F- mcrA ...use, dilute your antibiotic into your LB medium at 1:1,000. For example, to make 100 mL of LB/ampicillin...Delta-lacX74 recA1 araDelta139 D(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG tonA::Ptrc ccdA DB3.1 Invitrogen...(TetR) ∆(ara-leu) 7697 araD139 fhuA ∆lacX74 galK16 galE15 e14- Φ80dlacZ∆M15 recA1 relA1 endA1 nupG rpsL...Delta-lacX74 recA1 araD139 Delta(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG Antibiotics commonly used...

...can use Figure 1 to compare the genomes of these two family members.   Figure 1: Gamma-retrovirus...Retroviruses” is an umbrella term which encompasses 1.) the gamma-retroviruses — that have been developed...of retroviruses called the lentiviruses (like HIV-1) — that have been developed into lentiviral vectors...Academy of Sciences of the United States of America, 54(1), 137–144. https://doi.org/10.1073/PNAS.54.1.137. ...1997). Functional RT and IN incorporated into HIV-1 particles independently of the Gag/Pol precursor protein...RNA as template. Journal of Molecular Biology, 79(1), 163–183. https://doi.org/10.1016/0022-2836(73)90277...Aids Full-Length Human Immunodeficiency Virus Type 1 DNA Synthesis. Journal of Virology, 82(5), 2376. https...

... strand and a tyrosyl residue of topoisomerase I (1). If a 5′ hydroxyl group from a different DNA strand...Let’s break down the steps needed for TOPO cloning: 1. Create Your PCR Product: Design standard primers ...mind that Taq polymerase has an error rate of about 1 in 3,500 bases. Typically polymerases with proofreading...enzyme and Taq, with Taq used in an excess ratio of 10:1. Gel purify your PCR product and incubate it with ...you to see colonies within 8 hours. References: 1. Shuman S. "Recombination mediated by vaccinia virus...the topoisomerase site inserted into a beta-galactosidase cassette allowing a researcher to perform blue-white...

...how to use our deposit spreadsheet to complete Step 1. Steps 2 and 3 are usually very easy — our tech transfer...scientist, you will have the most involvement with Step 1, and, while data entry can be a chore, we’ve made ...Submission” and “Spreadsheet Submission” (see Figure 1). Please read our “How to Deposit Your Plasmids with...thorough guide to our online data entry process. Figure 1: Begin the deposit process by clicking “Online Submission...spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down options for you to choose...and are exceptions to our standard workflow.  Table 1: Guidelines for the data fields in the deposit spreadsheet...), M. musculus (mouse), R. norvegicus (rat), G. gallus (chicken), B. taurus (bovine), X. laevis (frog)...

... References 1. Casadaban M.J., Martinez-Arias A., Shapira S.K., Chou J. Beta-galactosidase gene fusions... fusions that retain their individual activities (1). Researchers also discovered the modular format of...systems could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown ...Although the original Y2H systems utilized the yeast Gal4 activator, bacterial LexA DBD and the lacZ reporter...

...near-perfect physical-to-infectious particle ratio of 1:1 (Zeltner et al., 2010). However, for recombinant ...preparations are genome-containing (Figure 1).    Figure 1: Electron micrograph of an Addgene AAV...reported a physical-to-infectious particle ratio of 50:1 (Zeltner et al., 2010). The specific infectivity of... lot (if we still have it available). References 1. Fagone, Paolo, et al. "Systemic errors in quantitative... Human Gene Therapy, Part B: Methods 23.1 (2011): 1-7. PubMed PMID: 22428975. PubMed Central PMCID: PMC3640491...8610.4. Hindson CM, Chevillet JR, Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL, Tewari M...

...protocols. A typical ExM protocol (Fig. 1) follows this structure: Step 1: fix (and maybe label) your sample...microscopy. Nat Biotechnol 1–12. https://doi.org/10.1038/s41587-022-01546-1  Mikhaylova M, Cloin BMC, ...https://doi.org/10.1038/s41592-018-0238-1  Klimas A, Gallagher BR, Wijesekara P, et al (2023) Magnify ...just adding water.)  Step 5: Image   Figure 1: The ExM process allows you to physically magnify ...

...is here to help. Contact us at [email protected] or +1 617-225-9000, or check out our MTA reference pages...s the heart and soul of Addgene’s mission), but legal complexities can arise when scientists are sharing...sharing space, we heard over and over again that the legal documents were one of the biggest hurdles for scientific... of not just the physical samples, but also the legal documentation that allows you to conduct your experiments...easy Even with all those benefits, we know that legal documents can get tricky. Here’s how Addgene helps...

...does not greatly alter the size of the tag.   Table 1: Common protein tags Tag Epitope Mass (kDa) Function... conditions FLAG DYKDDDD or DYKDDDDK or DYKDDDK 1 Affinity and Purification Good for antibody-based ...Function Notes CBP KRRWKKNFIAVSAANRFKKISSSGAL 4 Affinity and Purification Binding and elution steps use...eukaryotes TAP GRRIPGLINP WKRRWKKNFI AVSAANRFKK ISSSGALDYD IPTTASENLY FQGEFGLAQH DEAVDNKFNK EQQNAFYEIL HLPNLNEEQR...PILDEIADEY QGKLTVAKLN IDQNPGTAPK YGIRGIPTLL LFKNGEVAAT KVGALSKGQL KEFLDANLAG SGSGHMHHHH HHSSGLVPRG  12 Solubililty...

...modifications). Ready to get started?   Figure 1: Break free from the ball and chain!   Epigenetics...?  Dividing cells depend on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental to...interact with DNA independently of dCas9 targeting (Galonska 2018). This means that off-target loci – in neighboring... 301, 89–92. https://doi.org/10.1038/301089a0. Galonska C, Charlton J, Mattei AL et al (2018). Genome-...

...) vector (Figure 1). Everything else outside the ITRs gets left behind.    Figure 1: Recombinant AAV ...productivity but increases transgene expression. Sci Rep, 7(1), 5432. https://doi.org/10.1038/s41598-017-04054-4...Y., Nauwynck, L., Bonnin, D., Merten, O.-W., & Galibert, L. (2017). Impact of Inverted Terminal Repeat...

...give your research more visibility.    Figure 1: Illustration of SARS-CoV-2 from Philippa Steinberg...constructs include proteins like nonstructural proteins 1 to 16, spike proteins, envelope small proteins, and...Humana Press, pp 331–338 https://doi.org/10.1007/978-1-59745-181-9_23 Additional resources on the Addgene...backbones which drive expression using either TEF1 or GAL1 promoters. These vectors are intended for the high...

...discovered such as Kaede and PS-CFP2 (see Table 1). Table 1: Properties of Selected Irreversible Photoactivatable...) by mutating Threonine 203 to Histidine (T203H) (1). wtGFP normally contains a mixed population of neutral...in fluorescence when illuminated with blue light (1). Similar to PA-GFP, scientists later developed red...fixation conditions to preserve cellular structures (0.5-1% osmium tetroxide). These conditions destroy most ...with extraordinary molecular detail.   References 1.Patterson, George H., and Jennifer Lippincott-Schwartz...is believing." Annu. Rev. Biomed. Eng. 10 (2008): 1-38. PubMed PMID:18647110. Additional Resources on ...16980368. PubMed Central PMCID:PMC1635685. 12.Paez-Segala, Maria G., et al. "Fixation-resistant photoactivatable...

...within the mitochondrial matrix.     Figure 1: A chart with MoClo kit recommendations for different...Traditional Biology.” ACS Synthetic Biology, vol. 5, no. 1, American Chemical Society (ACS), 4 Nov. 2015, pp...., vol. 4, no. 9, American Chemical Society (ACS), 1 May 2015, pp. 975–86. Crossref, https://doi.org/10.1021...Yeast Mitochondria.” BMC Biotechnology, vol. 17, no. 1, Springer Science and Business Media LLC, 13 Nov. ...Editing in Plants.” BMC Plant Biology, vol. 20, no. 1, Springer Science and Business Media LLC, 23 Apr. ...Arabidopsis Mutants.” The Plant Journal, vol. 106, no. 1, Wiley, 25 Mar. 2021, pp. 8–22. Crossref, https://...Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media LLC, 12 July ...

...expression inducible by red light. pCph8 50552 pLtetO-1 Anhydrotetracycline Jeffrey Tabor Anhydrotetracycline...Plasmids placO with lambda-C1 operator Beta-galactosidase Ann Hochschild Bacterial two-hybrid assay using...Plasmids Sequence chosen by researcher Beta-galactosidase Keith Joung Transcription factor testing: Kit...

...these stabilized promoters a try. References 1. Segall-Shapiro, Thomas H., Eduardo D. Sontag, and Christopher...gene expression stable regardless of copy number (Segall-Shapiro et al., 2018). Designing stabilized promoters...constant. In the case of the stabilized promoters from Segall-Shapiro et al., they chose a TALE protein as the...replication, the stabilized promoters created by Segall-Shapiro et al. showed nearly no position-dependent...as cell division slows (Kittleson et al., 2011). Segall-Shapiro et al. showed that the stabilized promoters...number during the transition to stationary phase, Segall-Shapiro et al. placed a trans-acting RepA protein...genes whose proteins are needed sequentially. When Segall-Shapiro et al. placed the genes under the control...