... experimental design. How-to guide (Link opens in a new window) for the molecular design and testing of...interesting in implementing intersectional experimental design. The Deisseroth Lab maintains a Standard Operating...Gray Facilitate the Itch-Scratching Cycle via Descending Regulation. Neuron 101(1):45-59. PubMed (Link...

...using in mouse transgenics. Designing an Easi-CRISPR knock-in allele To design a knock-in allele at a given... PAM or guide mutation. Designing an Easi-CRISPR conditional allele The design for a floxed conditional...successfully used guides targeting the same or complementary strands, as well as guides facing towards or...How to design a conditional allele with Easi-CRISPR. As in the knock-in example, you’ll design your ...with your desired insertion feature (e.g., a fluorescent protein), you can amplify the desired region using... Figure 5: How to design Easi-CRISPR repair templates using Addgene plasmids. Design PCR primers to add...arduous. First, you’d construct a plasmid with your desired changes, long homology arms and a selection marker...

...recently described a rapid and robust protocol for constructing TALE repeat arrays of any desired sequence...Joung Lab REAL Assembly TALEN Kit , which also includes the four TALEN expression vectors. More detailed...

...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts To make targeted epigenetic...catalytically dead dCas9 to epigenetic modifiers. Design your gRNA to target a specific promoter or enhancer...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...

...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Catalytically dead dCas9...gene expression by interfering with transcription. Design your gRNA to target your gene of interest’s promoter...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...

...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Catalytically dead dCas9... can increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...

...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts CRISPR/Cas nickase mutants...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...Selectable Marker PI Publication Yeast ID Plasmid Description Gene/Insert Promoter Selectable Marker PI Publication...

...relaxed gRNA design parameters. To demonstrate the effectiveness of their library design, Moffat laboratory...-strand DNA (ssDNA) oligonucleotides (oligos), and thereby rapidly create desired allele combinations ... pCAG-T7-TALEN(Sangamo)-Destination series, pCAG-Golden-Gate-Esp3I-Destination Takashi Yamamoto pcDNA-TAL-NC2...vector, and besides being used for restoring strain prototrophy, can also aid in designing self-establishing...assay. In their recent publication, Kroeze et al. describe their enhanced TANGO arrestin recruitment assay...optimized, expression validated GPCR sequences to any desired backbone.  Kroeze, et al. Nat Struct Mol Biol....plasmids with the Cross-disciplinary Integration of Design Automation Research lab Moduclar Cloning (CIDAR...

...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Type VI CRISPR systems,...regardless of homology to the crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which require...encyclopedia of CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...

...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Fully functional CRISPR...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...or search the tables below for CRISPR plasmids designed to introduce a DSB. Plasmids are available for...

...Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have been designed for use in ...CRISPR plasmids that have been designed for use in zebrafish. CRISPR...

...clone in their desired DNA sequences. Gateway technology relies on the two reactions described below: The ...pLenti CMV Puro DEST (w118-1) or the doxycycline-inducible pLIX_403. The chosen attR destination vector will...between the entry clone and destination vector creates two products: the desires expression clone and a byproduct...produce the desired expression clone. This is possible thanks to the Gateway vectors’ design. They have...vector systems. With the appropriate entry and destination vectors, one can use Gateway to clone a gene ...generated entry clone and the attR sites of the destination vector. This reaction is catalyzed by the LR ...containing the ccdB gene is excised from the destination vector.  Once the BP and/or LR reactions are ...

...many different types of proteins. For a full description (including peak response spectra) of different...preparations of many optogenetics plasmids. ID Plasmid Description Gene/Insert Vector Type Promoter PI Publication...transcription", "Hahn", or other terms. ID Plasmid Description Gene/Insert Vector Type Promoter PI Publication...

...laboratory for engineering TAL effectors, including designed TALENs, and assembly methods for REAL, REAL-Fast... engineering TAL effector repeat arrays of any desired length: the REAL (Restriction Enzyme And Ligation...for FLASH assembly. Links to additional pages describing these reagents are provided below: Joung Lab ...

...proteomic interactions represent the framework that provides biological context. Assays monitoring impedance...sub-cloning into a wide range of Gateway-compatible pDEST vectors. ID Plasmid Gene/Insert Industry PI Pooled...solute carrier family of proteins. Library ID PI Description Human SLC Activation Library 132561 Superti-Furga...

....  These primers are typically ~70 nucleotides long (20 nucleotides that anneal to the DNA sequence of..., the substrate should be ~70-100 nucleotides long with the desired alteration(s) located in the center...genomes. 2) Designing ssDNA oligos that avoid MMR activation:  There are two ways to design oligos that...tube. The donor DNA substrate only requires ~50 nucleotides of homology to the target site for recombination... be explained in greater detail. Substrate DNA design and generation Expression of lambda red recombination...antibiotic resistance cassette.   Substrate DNA Design and Generation Whether you use a linear dsDNA or...insertions or deletions greater than approximately 20 nucleotides, while ssDNA substrate is best for point mutations...

...CoV-2), modification, or variant. ID Plasmid Description Mutations Industry PI Due to export control regulations...expressing mCherry and firefly luciferase. pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-Hygro - Lentiviral...Lentiviral vector expressing destabilized EGFP. Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing...

...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Two classes of base editors...protein (Mu- GAM). While many base editors are designed to work in a very narrow window proximal to the...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...

...list of Fragmid destination vectors currently available. Table 1: Fragmid destination vectors Organism...website, which breaks the design process down into four easy steps, provides the real value for the user...potential of them not working, or spend a few weeks designing and cloning the vectors you actually want. But... Fragmid is a modular vector assembly process designed for a quick, plug-and-play approach to cloning...et al., 2024). It reduces the amount of time to design and create a new vector to four days for an experienced...high school interns, were able to use Fragmid to design and clone their vectors in five days. Did you say...s modular assembly structure takes a lot of the design work out of the plasmid creation process. Each ...

...protein with many available isoforms and mutants provides a shared experience for the research community...site loop", or "subunit interface") ID Plasmid Description Gene/Insert Mutations Additional Resources MDH...Results in Student Learning Gains and Positive STEM Attitudes in CUREs. Callahan KP et al. CBE Life Sci Educ...