... both substantial merit and national importance; (2) The foreign national must be well positioned to advance...outstanding achievements in your specific academic field, (2) that you have three 3 years of research experience..., Casseus Law suggests a mix of experts including 2 dependents –people who have worked with you i.e. your... issues.  The permanent residence “green card” process is a necessary and often overlooked part of career...start planning for your visa and or green card process well before addressing your career alternatives... benefit from the applicant thinking about the process early, keeping good records, and planning for the...organizations do not have the April 1 H-1B queue process or quota like industry, these organizations can...

...PMID: 25386923. PubMed Central PMCID: PMC4227841. 2. Young, Carissa L., Zachary T. Britton, and Anne S...

...expected plasmid features.  Figure 2. Addgene’s basic QC process. Created with BioRender.com. Once...quality control (QC) processes to screen for and reduce these errors. These processes allow us to have a...developed a process for identifying, categorizing, and resolving a variety of QC issues. This process ensures...discrepancies. While Addgene’s QC process adds time to the deposit process, we believe this to be non-negotiable...research, these errors can affect experimental processes, results, and subsequent conclusions. Being vigilant... edit, and easy to share. However, the cloning process — including the propagation of plasmids in bacteria... [email protected] to get started! Addgene’s QC process Addgene prioritizes the sequence quality of the...

...either SARS-CoV-2, Adenovirus type 41 (AdV-41), or Adenovirus-Associated Virus type 2 (AAV2) – or some... gene expression: Implications for gene therapy. Proc Natl Acad Sci U S A, 95(19), 11377. https://doi....Undergoing Spontaneous Degeneration in Tissue Culture. Proc Soc Exp Biol Med. 84(3), 570–573. https://doi.org... was either 1) a physiological effect of SARS-CoV-2 toward destabilizing one’s natural immune response...response to Adenovirus (and/or to its partner, AAV) or 2) a sociological effect of the COVID-19 lockdowns causing...… Chiu, C. Y. (2023). Adeno-associated virus type 2 in US children with acute severe hepatitis. Nature...encode proteins essential to initiating the whole process – such as the viral DNA polymerase – it makes sense...

... “Ligand-Regulated Site-Specific Recombination.” Proc. Natl. Acad. Sci. USA 92(June): 5940–44. PubMed ...Using a Ligand-Dependent Chimeric Cre Recombinase.” Proc Natl Acad Sci U S A 92(15): 6991–95. PubMed PMID...FLP and Cre for additional control   Figure 2: Expression of a morphological marker (eYFP) and synaptic...Sternson. 2008. “A FLEX Switch Targets Channelrhodopsin-2 to Multiple Cell Types for Imaging and Long-Range ...PMID: 18614669. PubMed Central PMCID: PMC2593125. 2. Buchholz, F, P O Angrand, and A. Francis Stewart....The FLP Recombinase of the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently to DNA via a Phosphotyrosyl...

...genomes. Nern A, Pfeiffer BD, Svoboda K, Rubin GM. Proc Natl Acad Sci USA. 2011 Aug 23;108(34):14198-203...Allak B, van Drunen E, Kanaar R, Berns A, Jonkers J. Proc Natl Acad Sci USA. 2001 Jul 31; 98(16): 9209–9214...CHO) cells, as well as in cortical neurons in layer 2/3 of the mouse brain. Rubin’s work shows that the ...

...Addgene. 2. Ma, Hanhui, et al. “Multicolor CRISPR labeling of chromosomal loci in human cells.” Proc. Natl...two genes over time, at a range of distances from 2-75 Mbp. Labeling multiple, intrachromosomal loci Building...successfully resolved loci with physical map distances of 2 and 75 Mbp, with the calculated fluorescent distances...established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that they could evaluate the...dCas9 supplied by a separate vector. Figure 2: CRISPRainbow generates 6 unique colors and a white...

...topoisomerase I in Escherichia coli is sequence specific."  Proc Natl Acad Sci U S A.  1991 Nov 15;88(22):10104-8... the two DNA strands and releasing topoisomerase (2).  These days commercially available TOPO kits provide...Taq polymerase using your favorite PCR protocol.  2. Set Up TOPO Cloning Reaction: Mix together the PCR...PubMed PMID: 1658796. PubMed Central PMCID: PMC52876. 2. Novel approach to molecular cloning and polynucleotide...restriction enzymes or ligase, and requires no post-PCR procedures. Sounds easy right? The technique relies on the...insert, the magic of TOPO cloning happens. Basic procedure Let’s break down the steps needed for TOPO cloning... everything you need for the next step before proceeding. Pre-warming your antibiotic-containing plate...

...Reyes-Gutierrez P, Wolfe SA, Zhang S, Pederson T. Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7....resolved loci with physical map distances of 75 and 2 Mbp, with the calculated fluorescent distances correlating...established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that they could evaluate the...

...) by temperature-sensitive Sendai virus vectors. Proc Natl Acad Sci U S A, 2011. 108(34): p. 14234-9. ... temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci U S A, 2012. 109(27): p. E1848-57....revert back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then proliferate and redifferentiate... more common delivery methods include retrovirus [2], lentivirus [4], adenovirus [5], Sendai virus [6]...2006. 441(7097): p. 1061-7. PubMed PMID: 16810240. 2. Takahashi, K., et al., Induction of pluripotent stem... Pluripotent Stem Cells. Cell Stem Cell, 2015. 17(2): p. 178-94. PubMed PMID: 26189426. 15. Si-Tayeb, ...induced pluripotent stem cells. Stem Cell Rev, 2015. 11(2): p. 242-53. PubMed PMID: 25578634. PubMed Central...

...mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5547-51.... hybrid derived from the trp and lac promoters.” Proc Natl Acad Sci U S A. 80(1) (1983): 21-5. PubMed ...immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101/gad.2.6.718. PMID: 2843425...Targeted Gene Expression in Drosophila.” Genetics 186(2) (2010): 735-55. PubMed PMID: 20697123. PubMed Central... in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197-214. PubMed PMID: 22129153. Additional...over the TetO elements, allowing transcription to proceed (pJKR-H-tetR from the Church lab uses this system...Response Element (TRE)), allowing transcription to proceed. When tetracycline or one of its derivatives is...

...piggyBac transposase tools for genome engineering.,” Proc. Natl. Acad. Sci. U. S. A., vol. 110, no. 25, pp...provides a simple tool for conditional cell ablation.,” Proc. Natl. Acad. Sci. U. S. A., vol. 105, no. 38, pp...without leaving behind any accessory DNA sequences [2], [3]. Simply put, the piggyBac™ transposon containing...with a Footprint-Free™ genome edit.   Figure 2: Editing DNA using the piggyBac™ transposon selection...PMID: 26211958. PubMed Central PMCID: PMC4663986. 2.  A. M. Singh, V. V Adjan Steffey, T. Yeshi, and D...to transfect, select, and isolate clones. This procedure is cumbersome and inefficient, as it relies on...incorporated, and used during the differentiation process. Moreover, a splinkerette-PCR assay may be used...

...sensitize and kill antibiotic-resistant bacteria.” Proc Natl Acad Sci U S A. 9;112(23):7267-72. (2015). ...Citorik Plasmids here). In both cases, they used 1-2 CRISPR crRNAs targeting sequences found only in certain...resensitize antibiotic-resistant bacteria (Figure 2). The first phage introduced is lysogenic, integrating...machinery targeting 1) the given resistance gene and 2) a second lytic phage. Bacteria that receive this ...bacteria have previously flourished.   Figure 2: Two-phage strategies to resensitize bacteria to antibiotics...237163. Find plasmids from this paper at Addgene. 2. Bikard, David, et al. “Exploiting CRISPR-Cas nucleases...Most antibiotics target one of three essential processes: cell wall synthesis, DNA replication, or protein...

...CRISPR labeling of chromosomal loci in human cells.” Proc. Natl. Acad. Sci. USA 112(10) (2015): 3002-7. PubMed... to DFHBI in the presence of that metabolite (Fig.2). Spinach eventually evolved into Spinach2, a more...PMID: 25713381. PubMed Central PMCID: PMC4364232. 2. Baker, Monya. “RNA imaging in situ”. Nature Methods...emerging environmental pollutants.” Front Chem. 2014; 2: 41. PubMed PMID: 25019067. PubMed Central PMCID: ...biosensors.” TrAC Trends in Analytical Chemistry 27(2):108-117 · February 2008. 11. Svensen, Nina and Jaffrey...Filonov, Gregory S., et al. “In-gel imaging of RNA processing using broccoli reveals optimal aptamer expression...Aptamer-based assay for prion diseases diagnostic”. Procedia Environmental Sciences. Volume 12, Part B, 2012...

...expression and high cell-type specificity.  Strategy #3 (ProC): Create promoters by repeating mouse cell type ...retinal cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using phylogenetically conserved...would express Cre-mCherry and GFP.   Figure 2: Synthetic promoters can be paired for targeted expression...1345-1356. https://doi.org/10.1038/s41593-019-0431-2  Neff, E.P. An AAV library for retinal cell expression...

...identification.   Fig. 2: Gel image of Plasmid 61564: pBAMD1-2 (~4.7 kb) digested with SpeI and... higher multiple circular DNA forms in bacteria. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1406-13. doi... at ~3.6 kb and ~1.1 kb (lane 3). Uncut DNA (lane 2) has a band at <3 kb, which corresponds to the supercoiled...formation of plasmid multimers. Cell. 1976 Dec;9(4 PT 2):707-16. doi: 10.1016/0092-8674(76)90134-3. PMID: ...dimers in Escherichia coli K-12. Plasmid. 1989 Mar;21(2):138-41. doi: 10.1016/0147-619x(89)90057-7. PMID: ...Plasmids and Chromosomes. Microbiol Spectr. 2014 Oct;2(5). doi: 10.1128/microbiolspec.PLAS-0025-2014. PMID...Behavior of Plasmid DNA. Microbiol Spectr. 2015 Apr;3(2):10.1128/microbiolspec.PLAS-0036-2014. doi: 10.1128...

...mitigation with cross-over insensitive ATG-out vectors. Proc Natl Acad Sci USA 116:27001–27010. https://doi.org...one of lox2272 sites will now be sandwiched between 2 loxp sites and is excised by Cre (Figure 1). This ...flank the transgenes (non-homologous pairs) (Figure 2.). They found that almost all off-target expression...exercise caution when using them.    Figure 2. Schematic diagram showing the mechanism of spontaneous... Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse Brain. PLoS ONE 8:e76310... Pseudotyped with Viral Capsids from Serotypes 1, 2, and 5 Display Differential Efficiency and Cell Tropism...

...complete degradation of proteins in mammalian cells. Proc Natl Acad Sci U S A. 2012 Dec 4;109(49):E3350-7....required components are 1) a SMASh-tagged protein and 2) asunaprevir (small molecule). dTAG The degradation...gone through an additional round of evolution (AID-2) to reduce some of the leakiness associated with the...

...Recruitment of CRISPR-Cas systems by Tn7-like transposons. Proc Natl Acad Sci USA 114:E7358–E7366 . https://doi....et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although...cells of the plasmid after integration. Figure 2: (A) Streamlined VchINT expression constructs. (B)...genomic target site bound by QCascade (Fig. 1B). The process also creates a 5-bp target-site duplication (TSD...

...systematic studies of membrane protein interactions. Proc Natl Acad Sci U S A. 2004; 101(33):12242–12247. ...retain their individual activities when separated (2), and that DBD and TAs from different systems could... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ...