...guide RNA (gRNA) targeting your genomic site of interest. This is commonly performed by transfecting cells...complex with a targeting gRNA, to your cells of interest. Cas9 RNPs are capable of cleaving genomic targets...CRISPR components to their experimental systems of interest. Using this method can alleviate many issues that...Hempstead is a Senior Scientist at Addgene.  Andrew's interests include genome engineering and microbiology.   ...

...fifth repeat Nobel Prize winner in history. Interestingly, he also adds to the disproportionately large...allowing every student to work on any topic of interest. The result was a positive experience for the ...generate new knowledge that will be of genuine interest to other researchers in their field. Indeed, sharing...Duboue et al., 2022), as they increase student interest in scientific career paths while also broadening...

... interaction trap), genes encoding proteins of interest(POI’s) are fused to TAs and DBDs (see Figure 2a...those cells where binding between the proteins of interest has occurred. Although the original Y2H systems...way, proteins that interact with a molecule of interest (the ‘bait’) can be ‘fished out’ for further study... random genomic fragments from the organism of interest, or a defined cDNA library (e.g. the entire proteome...

...technical challenges depending on their protein of interest. After several decades of trying to address these... protein? It is not generally in a cell’s best interest to overexpress a protein. Energy and cellular ...many tags and fusion proteins to your protein of interest would eventually get ridiculous (you generally...Life Sciences. Addgene pages and blog posts of interest: Which Fluorescent Protein Should I Use? Plasmids...

...If you’re interested in studying immunology or subpopulations of cells, you’ll soon find yourself encountering...markers to identify your specific cell population of interest. The markers you use, and the order you use them...like in Figure 3 above, until the population of interest has been successfully identified and isolated..../doi.org/10.5500/wjt.v8.i1.23 Jhunjhunwala, S., Aresta-Dasilva, S., Tang, K., Alvarez, D., Webber, M.,...

...but there are few genetic tools currently for terrestrial frogs. Cnidarian-Dinoflagellate symbionts (Virginia...food. They have chemical pathways generating interesting drug-like compounds especially cardenolides which...traits with good genetic mapping morphology and interesting behaviors. There is diversity in the social bonds...from other plants and are unusual enough to be interesting biologically. They have economic importance as...

...inactivated native restriction/methylation system. This means the strain can neither restrict nor methylate...expression of T7 RNAP from the genome; does not restrict methylated DNA fhuA2 lacZ::T7 gene1 [lon] ompT...RNAP is then available to transcribe the gene of interest from a T7 promoter on the plasmid. Many commercial...

... ask questions like: “Where does my protein of interest live within a cell,” “Does this disease change...Blotting, but it allows you to visualize a target of interest within an intact cell, tissue, or, in some cases...IF, the primary antibody targets the antigen of interest but is not attached to a fluorescent molecule,...products that all claim to target your protein of interest. Thankfully, there are a number of websites compiling...

...is then used to detect the amplified region of interest. Cas13 uses a complementary crRNA sequence to ...based detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA per microliter Time...Resources Many thanks to our guest bloggers Shravanti Surest from Iowa State University. Shravanti Suresh is...Sashital Lab, Iowa State University. She is actively interested in science communication and enabling access ...

...selection to isolate plants with the phenotype of interest. Genome engineering, on the other hand, allows...tumefaciens as a vehicle to deliver your gene of interest into a target cell line or organism (Presented...for Golden Gate mediated cloning of your gRNA of interest. Figure 1: Simplified schematic of agrobacterium-mediated...can be used as a vehicle to deliver your gene of interest (GOI) into plant cells. A) The system typically...

...screens to identify genes involved in a phenotype of interest. This blog post will discuss differences between...target sites occur ~1 in every 8 nucleotides, restricting gRNAs to 5 – 65% of the protein coding region... the lab. Importantly, for any modification of interest, it would be unwise to make conclusions on the...experimental strategy is clear: for any gene of interest, one should require that multiple gRNAs of different...

...far away brain regions expressing your gene of interest. This can cause issues in tracing studies, where...Promoters The promoter controls how the gene of interest will be expressed; it can be ubiquitous, cell-...particles per volume required to express the gene of interest, in the cells being studied, at an adequate level...Kügler, S. (2017). A MicroRNA124 Target Sequence Restores Astrocyte Specificity of gfaABC1D-Driven Transgene...

...protein of interest, and a negative control sample that does not express the protein of interest....detection using antibodies specific to the protein of interest. This protocol describes the basic steps for lysing... Precast SDS-PAGE gel SDS-PAGE running buffer Prestained protein ladder Ethanol iBlot 2 PVDF Mini Stack...depending on the cellular location of the protein of interest. RIPA buffer is suitable for most proteins but...the samples into the gel. Load 5–10 µL of the prestained protein ladder. Place the lid on the tank and...sample type and cellular location of the protein of interest. You may need to try a variety of lysis buffers...

...confidently conclude the function of a gene of interest. Tools for RNAi Arguably the greatest advantage...required dsRNA complementary to your gene or genes of interest to trigger RNAi activation. A typical RNAi experiment...that contains sequence against your gene(s) of interest and the reporter. This approach greatly enhances...coupling silencing of APT and any gene target of interest, only organisms that are actively silencing to...

...comments. Additionally we’ve seen what topics are of interest to you, our readers. Below I’ve compiled a list... from the past year. A quick glance shows that interest in CRISPR continues to grow as the genome editing...community, so email us at [email protected] if you are interested. Addgene's Most Popular Posts of 2014 How...

...N-terminal tags, coding sequences of the gene of interest, C-terminal tags, plant terminators and resistance...component of Golden Gate and GreenGate are type IIS restriction endonucleases. They are like other endonucleases...their own lab: be very careful with the type IIS restriction endonuclease of choice, BsaI, which can be very...

...gene of interest in neurons. This ability to be paired with any Cre-controlled gene of interest makes pmSyn-BFP-Cre...may want to further control where your gene of interest is expressed.  A cell-type specific Cre can provide...

...two phosphatases, PTPN1 and PTPN2, which are of interest as cancer immunotherapy targets but are embryonic...s laboratory at Harvard Medical School and is interested in improving CD8+ T cell responses to cancer....generation of exhausted CD8+ T cell subpopulations and restrains tumor immunity. Nature Immunology 20, 1335–1347...

...expression. Cultures are then lysed and the protein of interest is purified using an affinity tag. Additional ...Determining the best way to express one’s protein of interest can save a lot of time later. Protein purification...C-terminal. Each set of plasmids uses the same restriction enzymes for cloning (see schematic below), enabling...

... to on Twitter? Prior to writing this post, my interest in Twitter was fleeting. I’ve had an account for...Unfortunately, it never clicked for me. Recently my interest was renewed in part due to FOMO  but mostly because... scientist? If you don’t use Twitter, are you interested in getting started? Reply in the comments below...