...Agrobacterium infection. At approximately 7 kb, pSiM24 is 2 and 6 kb smaller than commercially available pCAMBIA...for both transient and stable gene expression applications in plants. This smaller, easily customizable...Agrobacterium are termed binary vectors because they must replicate in two species: Agrobacterium and E. coli. Like.... coli. However, they also need to be able to replicate in Agrobacterium to ensure that they are maintained... Thus, binary vectors contain two origins of replication, one each for E. coli and Agrobacterium, along...bases each. By switching the E. coli origin of replication and codon-optimizing the Agrobacterium ORI, they...constitutively active M24 promoter has been modified with duplicated enhancer domains and characterized in studies...

...might otherwise be hard to identify. Figure 2: Users can cross-interrogate screens from the literature...   Figure 3: Each publication featured in BioGRID ORCS has a dedicated summary page with PubMed...favorite gene was described in a CRISPR screen publication. You see this mentioned in the results section...manually curated and annotated from more than 45 publications, beginning with the seminal research articles...screen curated in ORCS is linked to the original publication via its PubMed identifier, which can also be ...a list of individual screens curated for the publication. Clicking on the screen name takes users to a... supplemental data tables available with the publication are also provided for the user. This approach...

...Nat Methods. 2009 Feb 6(2):131-3.  PubMed. Find plasmids from this publication at Addgene. Subach OM...Since the first research applications of GFP were published in the 1990s, biologists have spent a lot...color was observed after excitation at 488 nm, indicating that spontaneous conversion did not occur. Tobacco... Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP photoconversion One concern...nmeth.1664. PubMed. Find plasmids from this publication at Addgene.   Addgene’s Fluorescent Protein...

...Genome Editing and The CRISPR Software Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed...job - we strive to also give you high-quality educational resources to help you do better research. CRISPR...design, sorting the plasmids we have available by category. Each plasmid page now has a helpful graphic to... newest page in the Transcriptional Regulation category already has five blue flame plasmids covering ...providing an avenue for temporary/reversible CRISPR modification. In addition to these subpages, our updated ... HypaCas9 Day 10: Transcriptional regulation: Truncated gRNAs for Regulating Gene Expression and CRISPR...

...E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference between arg1 and arg2 vesicles...Knowing where bacteria are located within their host is often key to understanding their role in both...reporters are not as well suited for imaging organs located deep in the body, like the gastrointestinal tract...lab recently discovered that gas vesicles also scatter sound waves and therefore produce signals in ultrasound... experiment to demonstrate the most exciting application for ARG’s: in vivo imaging. Specifically, arg1...

...posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing Mice. My first CRISPR mouse...is less predictable. In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA...interested in using CRISPR to develop therapeutic applications that better human health.     References Cong...

...Enhanced CRISPR-based DNA demethylation Figure 2: The components of Casilio-ME1. Image from Taghbalout...expression of specific Caulobacter genes using a catalytically dead Cas9 (dCas9) derived from Streptococcus...methylation-silenced genes. Read the paper in Nature Communications Find Casilio-ME plasmids If you have a new...

...CHO) cells, as well as in cortical neurons in layer 2/3 of the mouse brain. Rubin’s work shows that the ...Gerald Rubin’s lab sought to make complex genome modifications in Drosophila using multiple recombinases. To...offspring carried both the recombinase and GAL4, indicating that the recombinases were inherited normally...

...Joung 2016). Table 2: Unbiased Genotyping Options Technique Detection Limit Application Advantages Disadvantages...PMID: 23792628. PubMed Central PMCID: PMC3773023. 2. Vouillot, Léna, Aurore Thélie, and Nicolas Pollet...off-target sites across the genome and pinpoint the location of mismatches based on the sequences of the genome... Figure 1: Schematic representations of the indicated unbiased genotyping options. Figure reproduced...those relating to DNA structure (e.g. epigenetic modification, bulges) that may also impact off-target edits...cell-based methods are genome-wide, unbiased identification of double-strand breaks (DSBs) evaluated by... HTGTS Not reported Cell-based Identifies translocations Limited by chromatin configuration, produces...

...system. Nature Communications, 10(1), 1668. https://doi.org/10.1038/s41467-019-09656-2 More resources ...

...six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini...Bioinformatics 33.7 (2016): 1099-1100. PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to...the user. This approach is useful for several applications as will be discussed below. The software could...contain multiple gRNAs targeting all genes in the indicated species and were suggested to be the best gRNAs...

...their required qualifications and skill sets Taking the time to identify and research 2 – 3 potential...Effective verbal and written communication skills are key to publication and acquisition of funding. If...  Writing Communication Publicity Consulting internships Communication Teamwork Management...Models Communication Regulatory knowledge Networking Blogging Writing Communication Networking...scientists identify potential career pathways, education and mastering new skill sets must occur by doing... skills such as leadership, management, and communication. A great approach to fully understand and acquire... science class: Science Club for Girls) Communication to diverse audiences Organization Mentorship...

...project my team & I have poured our hearts into for 2 yrs) has 200K+ figures by 95K+ scientists 👨🏽‍🔬👩‍...software that let me create graphics that better communicated the science in my blog posts. This post highlights...free tools which will hopefully also help you communicate your science, whether it’s in presentations, ...number of drawing tools, so it’s harder to draw intricate figures, like a brain or a mouse. Vectr Vectr ...without the watermark, which are better suited for publications, if you sign up for a paid account.  I’m proud... free medical images organized into four main categories: anatomy and the human body, cellular biology...appropriate credit, provide a link to the license, and indicate if changes were made to the images. This requirement...

... Figure 2: Dense (c), intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, ... use the GAL4-UAS system along with precisely truncated attP sites to control recombination of the gene...of PhiC31. In the SPARC system, attP sites are located on either side of a stop cassette, upstream of ...amount of labeling, you can alter the attP site. Truncating the attP site upstream of the stop cassette results...population is directly correlated to the length of the truncated attP site. The three SPARC variants, based on ...lobe. Image from Isaacman-Beck et al., 2019. Applications for SPARC SPARC technology is versatile and ...dendrites of T5 neurons by placing the calcium indicator GCaMP6f in the effector position, as well as to...

...but also something about how science really works. 2)   Read science writing in magazines or newspapers... Foundation Round Table. In any field, good communication skills help when it comes to finding a job. ...an argument for taking the time to hone your communication skills. So, where can a graduate student or ...opportunities than ever before for scientists to communicate about their work and their interest in science...your area or at your institution; the news or communications office is a good place to start. It’s not unusual...about science writing as a career, the Science Communication Program at the University of California, Santa... your research stories. Consider practicing communicating your research by guest blogging here, just leave...

... temporal activation of a transgene. Figure 2: FLEX Cre-Lox system.  WPRE The Woodchuck Hepatitis...contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains the ... by the AAV. On a plasmid map, its sequence is located between the ITRs. It can be anything you like as...expression of your transgene to a specific species or location. Promoters can also drive different levels of ...sequences that can flank the transgene and are located between the ITRs. Transfer plasmids often have ... the expression of your transgene. The WPRE is located at the end of the AAV genome, just upstream of ...promotes RNA transcript longevity. The polyA is located at the end of the AAV genome, just upstream of ...

...access this tool, set up an IDT account. Figure 2: Addgene's Analyze Sequence tool.   DNA sequence...tackling these particular tasks, but they tend to be scattered about the internet. To help you keep your virtual...: Find sequences for plasmids assembled from publications and commercially available sources, as well ... as expression patterns, post-translational modifications and polymorphisms Codon optimization tool: To...making it ready to copy and paste into other applications Primer design tools Many of the above-listed... well as primer design for different cloning applications. Primer3 and Primer3Plus: For general primer...

...easily understood and consistent in design and layout. 2. Be concise in your writing  When writing a manuscript...of any scientist’s career. We use writing to communicate our findings and share them with the world. If...simply writing down your data doesn’t guarantee a publication. There is an art to composing manuscripts and...field of microbiology with multiple high-profile publications. My classmates consisted of graduate students...well as being critiqued added an extra level of application not present in the online class.   Four ways ...paper will be generated. After you complete a publication quality figure, produce the figure, its legend...

...Nature communications 9.1 (2018): 4425. PubMed PMID: 30356044. PubMed Central PMCID: PMC6200774.   2. Nielsen...published their results in a paper in Nature Communications announcing a new bioinformatics tool that can...at its sequence. MIT collaboration for gene classification This inquiry started when Neil Thompson, a ...have generated robust tools that have practical applications in monitoring the accidental or intentional ...the lab-of-origin of engineered DNA." Nature communications 9.1 (2018): 3135. PubMed PMID: 30087331. PubMed...

...method relies on the fluorescence ratio between the 2 fluorophores, rather than on the fluorescence intensity...be used to generate organoid cultures with a modification of interest. While the authors optimized the...mechanisms related to morphogenesis, cell fate specification, and human disease.     Sun, et al. bioRxiv...protoplasts, the enzymatic biosensor retains its catalytic activity.  As SLs are also known to trigger seed...parasitization of cereal crops. As a result, agricultural applications of these novel biosensors could have a major...