We narrowed to 610 results for: SCR
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TypeBlog PostPublishedJuly 15, 2014, 4:03 p.m....ligands NRs represent the largest family of transcription factors found in metazoans. The superfamily ...profiling datasets in to our data mining tool, Transcriptomine. We are currently nearing the completion of...transient transfection platform for assaying transcriptional activity, for example, was a key development...coregulators: Judges, juries and executioners of transcriptional regulation. Molecular Cell 27, 691-700. (PubMed...
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5 Steps to Writing Grant Proposals for Grad Students
TypeBlog PostPublishedFeb. 11, 2014, 12:13 p.m....includes overview information, key dates, funding description, research plan, award information, eligibility...aspects are understood. While reading the funding description, pay attention to the keywords used and think...grant applications restrict the space available to describe the proposed work. Thus, every single sentence...project are clearly stated and align with the description of the RFA and funding institution. Emphasizing... -
Educational Planarian Tools for Teaching Developmental Biology Techniques
TypeBlog PostPublishedMay 24, 2018, 1:20 p.m.... by T7 promoters to allow for bidirectional transcription to create dsRNA. Students can produce DNA templates...A recent publication [1] from the Sánchez lab describes the plasmids and protocols required for both WISH...the temporal and spatial localization of mRNA transcripts in tissue. This is useful for labeling specific... which are available through the deposit) are described in the Sánchez lab publication [1] so that the... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog PostPublishedJan. 5, 2021, 2:15 p.m.... protein to date By Justin Ng High-throughput screening methods, such as fluorescence-activated cell sorting...s lab developed a new form of FACS-based cell screening, Single-cell Phenotypic Observation and Tagging...plasmids here Reduce the scale of your CRISPR screens by using this lentiviral CRISPR human genome-wide...guides per construct and allows a reduced scale in screens without any reduction in performance compared to... -
A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog PostPublishedJuly 12, 2021, 1:15 p.m....loop-mediated isothermal amplification after reverse transcription (RT-LAMP) and a colorimetric readout that can... 45 minutes. The team deposited the reverse transcriptase, protease, and Bst polymerase constructs from...toolkit include ones for expressing MMLV reverse transcriptase and Taq polymerase that can be used in a one-step...Hsu PD (2021) Genome-wide, bidirectional CRISPR screens identify mucins as critical host factors modulating... -
Design Tips for Prime Editing
TypeBlog PostPublishedJan. 23, 2025, 2:15 p.m....on a fusion enzyme of Cas9 nickase + reverse transcriptase and a prime editing guide RNA (pegRNA). The ...) complementary to the genome and a reverse transcriptase (RT) template including the desired edit. ...., 2019). Test different lengths of reverse transcriptase template, starting with about 10–16 nucleotides...pegRNA primer binding site (PBS) and reverse transcriptase template (RTT). Created with BioRender.com. ... -
Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene
TypeBlog PostPublishedApril 6, 2021, 12:30 p.m....Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. Nat Protoc 12:828–863 . https:/...ask the depositing laboratory to review the discrepancies. If the depositing laboratory confirms that ... use a modified version of the Python program described by Joung et al. 2017, which we hope to make available... -
Tips for Titering Your Lentiviral Preps
TypeBlog PostPublishedMarch 15, 2016, 2:30 p.m....for viral titration consider using a reverse transcriptase mutant in a separate control reaction. This ...results. A packing vector deficient in reverse transcriptase will produce low levels of virus and can be ...into a target cell’s genome. This approach can discriminate multiple integration events, reducing the possibility... How Lentivirus is Used in CRISPR Genome-Wide Screening Learn How Lentivirus Has Been Used for Lineage... -
Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity
TypeBlog PostPublishedOct. 11, 2018, 12:30 p.m....suppression of background by antibiotic selection. Screening of transformants by colony PCR confirmed the insertion...obtained if any single component was omitted. Screening for successful Golden Gate Assembly All Golden...to faster assembly workflows that require less screening to confirm the accuracy of assembly. The fidelity... assemblies per plate using outgrowth volumes described in Figure 3, with calculated yields from entire... -
Transferable Skills: Negotiation
TypeBlog PostPublishedMarch 3, 2022, 2 p.m....how much data is necessary before submitting a manuscript. In fact, how many of you have negotiated over...over the order of authorship for a manuscript? And that’s only in your scientific life! I’m certain that...mate would have time to make comments on your manuscript. If you give her this slot, she’ll have extra...her deadline) to help you. Her feedback on your manuscript is more important to you than collecting your... -
Cloning Mammalian Cells with the Agarose Method
TypeBlog PostPublishedSept. 7, 2017, 12:17 p.m....up, most will eventually grow within a week. We screen the wells in groups as they become confluent, taking...important to have a functional assay in place to screen your clones. Depending on the protein, we have ...SH-SY5Y cells, as these cells did not seem to form discrete colonies but constantly migrated away from each...either initially, or using secondary biological screens. For example, for cancer researchers, if growth... -
Magnetic Control of Proteins: More than a Dream
TypeBlog PostPublishedSept. 24, 2024, 1:15 p.m....Maria were encouraged by previous reports that described magnetic responses of cryptochrome proteins or... and the team set up a directed evolution and screening experiment with three of the proteins that had...several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants were showing no... suggest she deprioritize the project, Maria’s screens of AsLOV2 variants showed some significant gains... -
Addgene's Cancer Collection Pages Connect the Research Community
TypeBlog PostPublishedSept. 6, 2016, 2:30 p.m....proteins involved, with the goal of being able to discriminately and effectively eliminate cancerous cells from...lines. Further, for each plasmid in the kit, the transcript most commonly found across all TCGA tumor samples...he or she can use to study it (as seen in the screenshot below). Currently, the page includes interactive... Tackling Cancers' Drug Resistance with a New Screening Kit Learn How the Clon Tracer Library Can Be Used... -
When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells
TypeBlog PostPublishedApril 19, 2016, 2:30 p.m....Green Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label ...world examples of these situations in cells are described below (See Figure 1). Figure 1: Each... not necessarily monomeric, even if they were described as monomeric in a publication. The gold standard... [2]. Some affinity assays used in the papers describing the FPs above including molecular sizing columns... -
Tips for Using BLAST to Verify Plasmids
TypeBlog PostPublishedMay 29, 2014, 1:29 p.m..... At Addgene, we use blastn to identify any discrepancies in Sanger sequences, such as mismatches, deletions...determine the potential effect of any nucleotide discrepancies. The blastp and blastx programs are optimized...matrix BLOSUM62, but you may want to check the description of the other matrices to see if another would...plasmids instead! All BLAST images are modified screen shots from the NCBI BLAST website. Additional Resources... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog PostPublishedSept. 6, 2018, 12:02 p.m....Cas9 expression The requirement for Cas9 transcription and translation delays editing What are Cas9...from Alex Schier’s Lab. gRNAs can be in vitro transcribed from ssDNA, which can be generated by commercial...targeted approach using RNPs has recently been described in vitro, with the hope of eventually using this...PMC5681539. Additional resources on the Addgene Blog Subscribe to CRISPR blog posts Learn about other CRISPR ... -
CRISPR 101: RNA Editing with Cas13
TypeBlog PostPublishedJuly 31, 2020, 12:30 p.m....C-to-U and A-to-I transcriptome-wide off target effects observed in whole-transcriptome RNA-sequencing, ...amount of off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced off-target...the RESCUE system’s ability to edit endogenous transcripts finding up to 42% editing efficiency. They note... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog PostPublishedSept. 28, 2017, 2:01 p.m....type of interest. Repeat rounds of infection and screening by PCR enrich for capsid variants with increased...of Chan et al, was neurons and astrocytes. Why screen for IV deliverable AAVs? Typically AAVs are directly...brain. Delivering the CREATE AAV library via IV screens for capsids that are able to cross this barrier...AAV as a tool for gene expression in mammals. Subscribe to Viral Vector Blog Posts Additional Resources... -
Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More
TypeBlog PostPublishedAug. 13, 2019, 12:58 p.m....sequenced genome also permits “whole-genome/transcriptome” studies, in which, for example, the activity...two revolutionary large-scale random mutagenesis screens produced ~1,500 zebrafish mutants that are still...deactivation tools for zebrafish Random mutagenesis screens provided a major toehold into understanding how...one-eyed pinhead mutant from the random mutagenesis screens. Microscopy-based lineage tracing can help illuminate... -
Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research
TypeBlog PostPublishedJuly 29, 2025, 1:15 p.m....an efficient gene activator when fused to a transcriptional activation domain (Karmakar et al., 2024). ... using your method of choice. Step 5: Colony Screening Use Primer 92F (5′-cattacgcaattggacgacaac-3′)...′) and Oligo 2 (specific to your target) to screen colonies via PCR. A 354-bp product confirms successful...insertion. Alternatively, use M13R and Oligo 2 to screen. A 667-bp product confirms successful insertion...