...) How many loci can Cas9 modify? To test the application of this system to gRNA multiplexing, Xie et al... indel frequency at both targets (35% and 76% biallelic), compared with only 44% and 66% indel frequency...were tested, they found that 50% of plants had biallelic mutations at all five loci. This highly efficient...multiplexing further illustrates the potential of PTGs. Applications of PTGs The major advantage of the PTG system... Mar 2. PubMed. Find the plasmids from this publication at Addgene Other Techniques for gRNA Multiplexing...May 15. PubMed.  Find the plasmids from this publication at Addgene Dimeric CRISPR RNA-guided FokI...Apr 25. PubMed.. Find the plasmids from this publication at Addgene ...

...should I choose? Writing a research paper for publication in a scientific journal requires several considerations...likely that researchers have enough information to replicate your experiments, making research more reproducible...the reproducibility and reliability of science publications. Peer reviewing To make sure your manuscript...happens before the manuscript is accepted for publication and is, in most cases, blind (meaning you don...revised before it can be published, this can delay publication and manuscripts may go through several rounds...a novel form of peer review that occurs post-publication and is completely open; the reviewer’s name and...scientific content of manuscripts submitted for publication. Holds a PhD in Cell Biology and a Masters degree...

...re used in a number of research and clinical applications. Most of the time, when someone speaks about...deeper penetration into a tissue, useful for applications like IHC, or be able to access an epitope folded...that would size-exclude an antibody. In imaging applications, smaller affinity reagents means higher resolution...antibody’s ‘Y’. They are often used in clinical applications. Fab fragments can be created in lab in situations...Shahied, 2004). They are often used for clinical applications, including imaging.  Diabodies are ~60 kDa proteins...antibodies are still an excellent option for many applications, and frankly, if ain't broke, don't fix it!  ...flexible in your approach to affinity reagent applications. Whatever affinity reagents you use, one thing...

...summarized in a recent Nature Medicine paper. In the publication, Bhang and coworkers used the ClonTracer library...propagated it, separated it into roughly identical replicate groups, and treated these groups with the same...below): 1. Resistant cells among the separate replicates would contain the same labels - i.e. they would...subpopulation 2. Resistant cells among the separate replicates would have different labels - i.e. they would...barcodes infect the target cell population at low multiplicity of infection (MOI) such that each individual... Looking at barcode abundances among multiple replicate populations, they found that similar barcodes ...populations were shared by at least one other replicate; prior to treatment, there are small (previously...

...scientists can mutate the transcription start site, splice sites, exons, and/or promoters to knock out lncRNA... and Bester 2021; Zibitt et al. 2021). Mutating splice sites is particularly effective as it can result...Transcription start site (TSS) deletion. (B) Mutations of splice sites. (C) Removal of an exon or a large genomic...it comes to targeting precision and practical application. If the CRISPR/Cas9 system is not effective for...lncRNAs with high specificity, particularly in applications requiring smaller delivery systems (Zibitt et...noncoding RNAs in human cells by Cas9 targeting of splice sites. Nat Biotechnol. Published online November...expression by lncRNA promoters, transcription, and splicing. Nature. 2016;539(7629):452. doi:10.1038/nature20149...

..., its dimeric nature can interfere with some applications. Several labs have been working to produce monomeric...Image reused from Ando et al. 2023, under CC-BY license.   The Miyawaki lab generated a series of ... for molecular fusion and membrane-targeting applications. Nature Methods, 1–9. https://doi.org/10.1038...Chuapoco, Flytzanis, Goeden, et al. 2023, under CC-BY license.   Find the AAV.CAP-Mac plasmid here!   Chuapoco...rats. Figure from Chen et al (2023) under CC-BY license.   Find AAV1-X1 plasmid here! Chen, X., et ...proven itself a good option for essentially all applications. Addgene’s plasmid collection includes over ...clone (M2) has been widely used in a variety of applications. Find anti-DYKDDDDK recombinant antibodies here...

...the best, or even which one will work for your application? Like many scientists, I have spent hours sifting...produced from a new set of animals), and wrong application (different experiments/conditions can change ...differently over time), compatible with the desired applications (such as immunohistochemistry (IHC) or western... members (like me!) are dedicated to creating a public and accessible database for low-cost, high-quality...positive, it will be released through the website for public reference. The Gift of Giving As an open-source...for goldilocks antibodies and, alongside that, a publicly available 2D/3D mouse atlas from antibodies that...resolving the antibody crisis. The mAb3D Atlas is a public service to the science community, and I am excited...

...Co-Ordinating Centre for Public Engagement (NCCPE) provides on-site courses in public engagement for institutions...tips on using social media as a scientist here. 2) Public engagement - Participate in science festivals and... and outreach If you are comfortable talking in public, volunteer at science festivals. Or engage with...performance. When your institute opens its doors to the public, for example at an anniversary or for Girls’ Day...for 10 busy weeks at media outlets like National Public Radio, WIRED, Scientific American or National Geographic...projects Lastly, if you'd like to directly involve the public in your research projects, you can set up a citizen...

...Shakespeare for the first time. The discovery and application of CRISPR-Cas9 kicked off over a decade of frenzied...to subtype to variant. Despite the seemingly complicated steps to arrive at this hierarchy, the main aspects... Class to type to subtype to variant (shown in italics). Created with BioRender.com.   Class 1 versus...action power of Cas3. Cas3 functions as both a helicase and a nuclease, unwinding and cutting up DNA after...nuclease effector. However, IV-C does in fact have a helicase domain due to a gene that resembles cas10. All... use in mammalian systems, with the landmark publication in 2013. Since that point, it has become the ...mechanisms in type I-C CRISPR-Cas3 for genome-editing applications. Molecular Cell, 84(3), 463-475.e5. https://...

...valuable to scientists, there are some potential complications that must be considered when using this method...alternative approach, which avoids many of these complications, is to directly deliver a ribonucleoprotein ...used for most of the current genome engineering applications of CRISPR including: generating single or multi-gene...pathways. This makes Cas9 RNPs useful for CRISPR applications where limited expression of Cas9 is required...protocols outlining these steps have been made publicly available by both the Jacob Corn and Alex Schier... of eventually using this method for in vivo applications. This technique utilizes Cas9 proteins harboring...CRISPR experiments can reduce many potential complications, there are additional factors to take into account...

... right AAV for your system or to successfully replicate published works. Of particular importance to these... focus on getting this information out into the public where it can be utilized by others. Addgene’s AAV...citable DOIs through DataCite before being made publicly available through the Data Hub. These minimum ...performance. Yet, most of the technical data needed to replicate and build on others’ results is available not ...information provided in openly accessible forms like publications. Why not get this data out there?  The Addgene...your data - negative AND positive - out into the public eye where it can benefit the community. We are ...Freedom: The Expansion of Data Sharing in Research Publications...

...performs well in many different antibody-based applications.  IPI’s recombinant version is a fusion of mouse... reused from Anderson et al., 2024 under CC BY license.    Find Anti-V5 [IPI-SV5-Pk1] Antibody here...adapted from Kroupova et al., 2024 under CC BY license. This construct can be easily expressed and...Image reused from Cao et al., 2024 under CC-BY license.  These optimized fluorescent proteins will...cellular processes in C. elegans: mScarlet3 for applications needing intense brightness (such as low-expression...first use of CRISPR/Cas9 in M. abscessus. The simplicity and efficiency of this dual-plasmid approach ...Image reused from Neo et al., 2024 under CC-BY license.   Find plasmids for CRISPR/Cas9 in M. abscessus...

...activity of CcdB. Applications of ccdB in plasmid cloning Perhaps the most widely used application of CcdB is...used as biological tools for a wide range of applications in molecular biology or in vivo studies. Let...types of plasmid addiction systems and their applications. The discovery of toxin-antitoxin systems Toxin-antitoxin...CcdA (antitoxin) where CcdB inferes with DNA replication by binding the GyrA subunit of DNA gyrase and...construct. Other toxin-antitoxin systems and their applications MazE/MazF Aside from CcdB/CcdA, there are many...Kulakauskas, Saulius, Arvydas Lubys, and S. Dusko Ehrlich. "DNA restriction-modification systems mediate ...

...to dramatically increase the number of CRISPR applications. The first non-Cas9 CRISPR protein adapted for...doi.org/10.1038/nbt.3609 Find plasmids from this publication at Addgene. Kim E, Koo T, Park SW, Kim D, Kim...10.1038/ncomms14500 Find plasmids from this publication at Addgene. Kleinstiver BP, Prew MS, Tsai SQ...10.1038/nature14592 Find plasmids from this publication at Addgene. Kleinstiver BP, Tsai SQ, Prew MS...doi.org/10.1038/nbt.3620 Find plasmids from this publication at Addgene. Hu JH, Miller SM, Geurts MH, Tang...10.1038/nature14299 Find plasmids from this publication at Addgene. Walton RT, Christie KA, Whittaker.../j.cell.2015.09.038 Find plasmids from this publication at Addgene. Additional Resources on the Addgene...

... citations for recent articles. Of course, a publication in Science or Nature is always nice, but in the... lesser-known journal as it is in a top-tier publication. Accessibility is surely also behind the increase...increase in citations for older publications. (As an interesting aside, Nature recently reported that the...in a solution.) But are citations and journal publications really the best way to measure impact? As that...and what it’s made of. Increasingly, journal publications alone aren’t enough. Scientists are sharing ...consider and quantify the impact of scientific publications. Ventures like Impact Story seek to find ways...

...decades. Widely used across various research applications, they have become indispensable tools for manipulating...-lasting changes in cells. However, for some applications, integration might be considered a double-edged... is distributed across multiple plasmids. For simplicity, we’ll use the second-generation system as an...dividing cells because they lack an origin of replication (Banasik and McCray, 2010). However, IDLV genomes... certain types of clinical or basic research applications. Considerations for IDLV use If you’re reading...Integrase-defective lentiviral vectors: progress and applications. Gene Therapy, 17(2), 150–157. https://doi.org...lentiviral vector: prospects for safe clinical applications. PeerJ, 10, e13704. https://doi.org/10.7717/...

...current CRISPR research focuses on its potential applications for human medicine (Waltz, 2016), the potential...selected the correct CRISPR components for your application, it is time to deliver these components to your...detergent, Polyethylene Glycol (PEG), although the application of this approach is limited to protoplast cells...find many plasmids for a wide variety of CRISPR applications in plants available through Addgene. In addition..., we highly recommend reading the associated publications or protocols to get the most out of the plasmid...N (2016) The CRISPR/Cas Genome-Editing Tool: Application in Improvement of Crops. Front Plant Sci 7: ....10.1073/pnas.1818290116 Zhu H, Li C, Gao C (2020) Applications of CRISPR–Cas in agriculture and plant biotechnology...

... February, the Office of Science and Technology Policy issued a memorandum calling on Federal agencies...in annual R&D expenditures to support increased public access to the results of federally-funded research...an organization called Free the Data launched a public database to gather the results of Myriad's genetic...support for open access to peer-reviewed journal publications, although many don’t necessarily publish in ...gave preference to journal access granted via a public repository or authors’ own web sites. 9)      The...to share science. Image credit: Judy Breck, via flickr  ...

... ReFigure. Reading and exploration including replications and experiments resulting in “negative data”...small selection of data found in full-length publications. As early career researchers, we wondered how...performance as in this antibody ReFigure Analyzing replications as in this ReFigure about whether Natronobacterium...reagent performs well, or you might publish a replication experiment that you conducted as part of setting...papers as they are not included in traditional publications. By submitting your data to a repository (Figshare...-registration, preprints, data, software, micropublications) in grant proposals. ReFigures are one such...

...Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS...tet-systems-legacy-products/tet-on-and-tet-off-2nd-generation Mullick, A., Xu, Y., Warren, R. et al. The cumate gene-... PMCID: PMC2942869. Find plasmids from this publication at Addgene. del Valle Rodriguez, Alberto, Dominic... PMCID: PMC2883883. Find plasmids from this publication at Addgene. Wei, Xing, Christopher J. Potter... PMCID: PMC3846601. Find plasmids from this publication at Addgene. Riabinina, et al. “Improved and ... PMCID: PMC4344399. Find plasmids from this publication at Addgene. Riabinina O, Potter CJ. The Q-System...