...Featured article from Journal of Visualized Experiments (JoVE) about genomic deletions in mammalian cell...video reprint from the Journal of Visualized Experiments (JoVE). The video publication by Stuart Orkin's...allows genetic perturbation of genes and genetic elements. Here we present a simple methodology for CRISPR...loss-of-function, and utility for the study of non-coding elements. This approach can be used for efficient loss-...loss-of-function studies of genes and genetic elements in mammalian cell lines. Protocol For referenced Tables...possible deletion strategies for genes and non-coding elements. For creating a gene knockout, two sgRNA located...6.3 . Assemble a 20 μl PCR with the following components: 10 μl 2x PCR mix, 0.5 μl forward primer (10 ...

... collections and resources are up-to-date with reagents and information you need to use CRISPR in your... for a variety of organisms, including mammals, plants, bacteria, yeast, Drosophila, zebrafish, C. elegans...of plasmids that are used together in CRISPR experiments. CRISPR Pooled Libraries : Find CRISPR libraries...knockout, activation, inhibition, and barcoding experiments. Browse CRISPR Plasmids by Function Genome Engineering.... Insert NEW CRISPR transposases insert large fragments of DNA. Transcriptional Regulation Activate dCas9...

...pseudovirus, where the structural and functional components of one virus (such as HIV, murine leukemia virus...information. Plasmids Expressing Spike and Spike Variants Full-length wildtype SARS-CoV-2 spike protein ...D614G change which is present in many circulating variants. The table below lists expression plasmids that...Other Resources Protocols Bloom lab - Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV...Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants Addgene COVID-19 Resources and Collections COVID...

...Purchase and the applicable material transfer agreements (MTAs) and limited use label licenses (LULLs)...10 business days in order to proceed with any adjustments to the original order. Turnaround Time Production...10 business days in order to proceed with any adjustments to the original order. Experimental Disclaimer...Disclaimer Due to the novel nature of experiments carried out with our materials, Addgene cannot guarantee the...assists importation by including thorough shipping documents and following import procedure, it is the responsibility...

...Cre-Lox system and how it can be used for genetic rearrangements and recombination. Use Addgene's curated tables...tables to find Cre-lox plasmids for your experiments....The schematic below shows the three types of rearrangements: inversion, deletion and translocation. Inversion...recombinase is split in half into N and C terminal Cre fragments (NCre and CCre, respectively) and placed under...Eisenhoffer 131782 pME-CreLite CreLite system components, PhyBdeltaCreC and PIF6CreN, in middle entry ...Eisenhoffer 131785 pAAV-CreLite CreLite system components, PhyBdeltaCreC and PIF6CreN, in AAV donor/transfer...Eisenhoffer 131786 pLenti-CreLite CreLite system components, PhyBdeltaCreC and PIF6CreN, in lentiviral vector...

...choosing what plasmid backbone to use, you have many elements to consider. Here is a guide to Addgene's... purification or localization experiments Yeast GAL4, PGK, ADH1, ADE2, TRP1 Gateway destination... pUAST vector containing both P-elements and attB site for high expression... transient expression is sufficient for some experiments, scientists often want to create stable ...basal activity shRNA/RNAi For gene silencing experiments pLKO.1 - TRC cloning vector ...

...generates “clean” homozygous mutants with no co-integrated markers or footprints, and can be scaled up for...This CRISPR/Cas plasmid-system consists of two components: A vector containing Cas9 and a validated gRNA...More details on how to use this plasmid in your experiments can be found below and in the supplemental materials... as homology arms. The system consists of two components: An all-in-one CRISPR/Cas9 vector based on the...

...) Ectodomain and RBD mutants that can be used to identify antibody escape mutants and for comprehensive...Libraries - Libraries of Spike (S) Ectodomain and RBD mutants (Timothy Whitehead). SARS-CoV-2 Spike Receptor ... Coronavirus Resource Hub COVID-19 SARS-CoV-2 preprints from medRxiv and bioRxiv Elsevier's Novel Coronavirus...differentially and only proteolysis by TMPRSS2 augments entry driven by the severe acute respiratory syndrome...

... launched in 2008 to drive the development of treatments and cures for Rett Syndrome and related MECP2...syndrome is a neurodevelopmental disorder that presents in early toddlerhood primarily affecting females...skills, notably loss of acquired purposeful hand movements and regression of speech. In contrast to the sustained...there is wide variability in symptoms even within patients with the same mutation, the location and type ...2007. Correlation between clinical severity in patients with Rett syndrome with a p.R168X or p.T158M MECP2...

...intron-lacZ/NLS Ple144 NTSR1 pEMS1137 EGFP/cre/NLS Ple144 NTSR1 pEMS1394 EGFP/NLS Ple146 NTSR1 pEMS1396 EGFP/NLS.../NLS Ple146 NTSR1 pEMS1617 intron-lacZ/NLS Ple147 NTSR1 pEMS1397 EGFP/NLS Ple148 OLIG1 pEMS1169 EGFP/NLS...

...systems, allowing for the integration of large fragments of DNA at specific locations. The Cas effector...recruiting the transposase (comprised of multiple components) to target locations in the genome defined by...for a ntiviral de fense), and four transposase components (TnsA, TnsB, TnsC, and TniQ). Type I CASTs can...V-K Cas effector, Cas12k, and three transposase components (TnsB, TnsC, and TniQ). Unlike type I, type V-K...

...TALEN Kits TALengineering Reagents Joung Lab TAL Effector Engineering Reagents You may also like... Keith...Reagents from the Keith Joung laboratory for engineering TAL effectors, including designed TALENs, and...assembly. Links to additional pages describing these reagents are provided below: Joung Lab REAL Assembly TALEN...

... with multiple Cas9 variants including high fidelity Cas9’s. Further advancements have been made by optimizing...systems create a wide spectrum of single-nucleotide variants (somatic hypermutation) in a wider editing window... for expression in mammalian systems, bacteria, plants, yeast, and zebrafish. Mammalian ID Plasmid Gene...

...leaders providing research opportunities to their students and other members of the MCC community. They support...courses, at any type of institution, and at different points in their careers to participate in the community...single protein with many available isoforms and mutants provides a shared experience for the research community...PMID: 35921892 (Link opens in a new window) Acknowledgments The MCC and plasmid construction was funded...

...Sarcomeric thick filaments 114414 MYL2-mEGFP AICSDP-49 mEGFP MLC-2v Sarcomeric thick filaments 124606 SOX2-...87425 ACTB-mEGFP AICSDP-15 mEGFP Beta-actin Actin filaments 87426 SEC61B-mEGFP AICSDP-7 mEGFP Sec61 beta Endoplasmic...Troponin I, slow skeletal type Sarcomeric thin filaments 114412 TTN-mEGFP AICSDP-45 mEGFP Titin Sarcomeres...

...mammalian cell types. AAV is used in a wide range of experiments, both in vitro and in vivo. It has low immunogenicity...AAV Packaged on Request, scientists can start experiments as soon as they receive their AAV prep from Addgene... processes to make sure it is ready for your experiments. Shipping 2–7 days We will email you with tracking...Once you start getting results from your AAV experiments, remember to share your protocols and results...

...interest 48807 8xLexA-binding elements/pMD20 iChIP plasmid, LexA-binding elements 48874 3xFNLDD/pCMV-7.1 iChIP... Efficient sequence-specific isolation of DNA fragments and chromatin by in vitro enChIP technology using.... PubMed . Direct identification of insulator components by insertional chromatin immunoprecipitation....

...specific QC experiments performed varies for each viral lot. To learn which specific QC experiments were performed...ddPCR) using primers and probes targeting the ITR elements and an internal control of known titer (protocol...viral genome and the absence of unexpected DNA contaminants. The data is also used to determine recombination...

...effect on the coding sequence, functional noncoding elements and off-target sites. This algorithm allows for...and allows the direct comparison of individual experiments. PubMed PMID 27404874 . MAGeCK Model-based Analysis...high-throughput design and analysis of CRISPR + HDR experiments. For more information, see the Addgene blog post... for the purpose of designing and evaluating experiments. Green Listed - a CRISPR Screen Tool Design gRNAs...

...these ORF fragments (which have become exons) determines the logical expression requirements of Cre and...References The development of powerful molecular reagents for biology have propelled our understanding of... for the molecular design and testing of novel INTSRECT plasmids for groups interested in producing their...