We narrowed to 976 results for: nin

...sequence of the plasmid, we examine three things: Aligning the NGS result to a reference sequence to confirm...backbone elements.   Confirming the gene/insert by aligning to NCBI entry or using BLAST.   Confirming tags.... We’ll break down each of these steps below. Aligning the sequence The first step we take when performing...publicly available reference sequence or plasmids containing genes with many modifications. We look for point...-Cas9 knockout and transcriptional activation screening. Nat Protoc 12:828–863 . https://doi.org/10.1038...

...treatment strategies for these disorders have involved turning on fetal hemoglobin to compensate for defects in...approach, editing patient cells ex vivo and then returning them to the patient. The first human trial involving...many, many pieces. Locus Biosciences is currently running this trial, with more updates expected in the coming...considerations for all CRISPR therapies, but especially concerning when using a systemic approach. Regarding effectiveness...difference, then the treatment will not work in a meaningful way. Delivery, especially for systemic approaches...

...lesson plans that are made freely available online; running professional development (PD) workshops for teachers...organizing Congressional briefings in Washington DC; convening meetings of experts and leaders from academia,...number of different opportunities to broaden my training beyond the biological sciences, as well as to ...students. One of these organizations was pgEd. After obtaining my PhD and working in digital academic publishing...festivals, and getting experience in either giving or running these events will be important both in preparing...

...produced in E1-containing mammalian cell lines such as HEK293 cells. 1st generation AdV cloning capacity is.... 2nd Gen AdVs were designed to have increased cloning capacity, decreased  potential for the generation...deleted in 2nd generation AdVs, thereby increasing cloning capacity to ~12 Kb. The ever popular pAdEasy system... interest for gene therapy due their increased cloning capacity (up to 36 kb!), long-term transgene expression...

..., Terns MP, White MF, Yakunin AF, Garrett RA, van der Oost J, Backofen R, Koonin EV. An updated evolutionary...transfer, analogous to traditional restriction enzyme cloning. Sticky-end mediated gene transfer would be particularly...homology-directed repair (HDR) over non-homologous endjoining (NHEJ). Cas9-mediated NHEJ usually destroys the...Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F. Cpf1 Is a Single RNA-Guided Endonuclease...

... antibodies may have similar emission spectra, meaning they will emit fluorescent light at similar wavelengths...record the fluorescence for both at similar levels, meaning that your populations will ‘look’ similar on your...compensate in the top right corner of a flow plot, meaning that it should be double-positive for the two markers... what two markers your fluorescent markers are staining for. If a cell is positive for both markers, it...compensation values later. This can be helpful as you're learning! Ashlyn has a research background in CD8+ T cells...

...collection of CRISPR tools has grown steadily, gaining approximately 2,000 plasmids/year from 2017 to ...to 2023. This growth is supported by new labs joining the Addgene CRISPR community, with approximately ...wanted to know, so we classified CRISPR plasmids containing a nuclease into three functional groups: nucleases...grown to 17,560 CRISPR plasmids. That’s almost a ninety-fold increase in size! From this collection, we...educational resources help scientists navigate that learning curve. As of 2023, our CRISPR guide has been viewed...

...naked eye. This makes cloning quite a bit easier since you can see if your cloning experiment worked just...methods. While techniques such as  blue-white screening require X-gal, an expensive exogenously-added ...chromoproteins don’t require other substrates for screening. Thus chromoproteins can be used as markers in... between chromoproteins. In some cases, cells containing high-copy plasmid-borne chromoproteins lost color...

... of these resources as I could. Networking and learning from the careers of others were key to creating...productive partnership CINSO has had. Volunteer! The turning point in my graduate experience, the point at which...Dinosphere” almost every week. I gained experience explaining science to visitors of all ages, learned a lot...lessons they have learned during their scientific training and wrote about them in a blog post on Born to...to Science: Life Lessons from Scientific Training More and more voices of encouragement emphasize the...

...delivery methods.   General considerations for planning your CRISPR experiment As with any experiment,... factors that need to be considered during the planning process. For CRISPR experiments, the following...viral delivery. Cas9 mRNA and gRNA Plasmids containing gRNA and Cas9 are used in in vitro transcription...as well as genome repair by non-homologous end joining (NHEJ) or HDR. EGFP validation of sgRNAs: Can evaluate...evaluate component delivery and sgRNA efficacy by cloning in genome target sequence into EGFP reporter. Target...

... direct RNA-guided transposition. By natively combining the DNA integration capabilities of transposases...with Type I-F INTEGRATE using a single plasmid containing a multi-spacer CRISPR array. Workflow and...s41586-019-1323-z Peters JE, Makarova KS, Shmakov S, Koonin EV (2017) Recruitment of CRISPR-Cas systems by ...Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin EV, Zhang F (2019) RNA-guided DNA insertion with...science.aax9181 Strecker J, Ladha A, Makarova KS, Koonin EV, Zhang F (2020) Response to Comment on “RNA-...

...detection had over 1,000 authors. The chapter on deep learning addresses the dilemma of creating a technology... “The most obvious future consequence of the undermining of the scientific process and the marginalization... the summer of 2020 that the consequences of undermining science are even more immediate and dire than...himself has written entire books about optogenetics. Defining science and summarizing its past, present, and...task to set oneself. As a scientist who enjoys learning a little bit about many different things (part...

...standard in their bicinchoninic acid assay (BCA) to normalize samples before running a western blot. Standard...provide some general considerations for scientists planning to use them in their assays.  What is a standard... above 0.95.    Additional considerations When running assays that rely on a standard curve, it is critical...program. Summary Few things in the lab are more disheartening than seeing a potentially exciting result quashed...

...plasmid cloning blog posts Resources on Addgene.org Find protocols for molecular biology and cloning Read... use a phenol chloroform extraction to remove remaining proteins and RNase. Finally, DNA is precipitated...into this mixture to absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids..., as detected by qPCR, but this did not reduce cloning efficiencies.  When compared to fresh columns, ...

...discretionary money. They also usually have a good event-planning base and will know how to reserve space, time ...themselves as “scientists.” We had some accountants running our math tables!  Be realistic about what you...you can teach  5-10 year olds in a single evening. Grades K-5 cover a broad range of attention spans, skill... simple. I honed the complexity of the talk by running it by the teachers and sharing some slides on Twitter...Even the most ambitious teachers, who work hard maintaining a current, dynamic curriculum, would be hard-...

...identifies nine enemies that stand in the way of creative thinking. I won’t go through all nine here, but...found that many researchers feel they lack formal training in a variety of transferable skills. At Addgene...absolutely a transferable skill developed during PhD training. Creativity is required to develop new projects...’re stuck drawing stick figures, remember your training as a scientist has been building your creativity...

... genome. These off-target sequences may be unintentionally mutated along with the desired target locus...sequences when used in place of wild-type SpCas9. The remaining variants (VQR, EQR, and VRER) recognize novel ...Staphylococcus aureus (Sa) NGRRT or NGRRN Neisseria meningitidis (Nm or Nme) NNNNGATT Campylobacter jejuni...Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F (2015) In vivo genome editing...Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F (2015) Cpf1 Is a Single RNA-Guided ...

...checkpoint.  The plasmid is self-transmissible, meaning that the plasmid is conjugative and carries the...inducibility (Toh et al. 2023), and additional cloning sites (Keen 1988).  Find broad host range recombinant... Origin of the Broad Host Range Plasmid RK2: Positioning of Various Motifs is Critical for Initiation ...1988). Improved broad-host-range plasmids for DNA cloning in Gram-negative bacteria. Gene, 70(1), 191–197...R. (1988). Chapter 15—Broad-Host-Range Plasmid Cloning Vectors for Gram-Negative Bacteria. In R. L. Rodriguez...

...HypaCas9, are great choices. Check out our SpCas9 containing plasmids, optimized for a variety of species....critical enzymatic residues has been converted to an alanine (D10A or H840A), changing its double-strand cut...available. This system utilizes an RNA template containing the desired edit adjoined to the sgRNA, a reverse....   Ok, I have a Cas9! What do i do next? Designing or selecting a sgRNA Addgene may carry a validated...expression systems Addgene carries a variety of Cas9 containing plasmids that have been optimized for expression...

... shift to remote learning due to COVID-19. We had many protocols for plasmid cloning and working with ...details about personal protective equipment to learning about how to use a tissue culture hood and other...on our Protocols page and YouTube page.  Happy learning, and welcome to the lab bench, new scientists!...