We narrowed to 411 results for: genome
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TypeBlog PostPublishedApril 30, 2024, 1:15 p.m....twist is that the typical genetic cargo — the viral genome — is swapped, at least partially, for a nucleic...other cells, but since the majority of the viral genome has been swapped for alternative genetic cargo,...nucleic acid cargo to be integrated into the host genome? Expressed episomally long-term? Expressed short-term...For example, some viruses integrate into the host genome (lentivirus and retrovirus) while others are non-integrating...
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Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein
TypeBlog PostPublishedJune 28, 2018, 3:01 p.m....coupled to high-throughput sequencing to identify genome-wide binding patterns of the target protein. Both...Transcript ID(s) of your favourite gene(s) from Ensembl genome browser: https://www.ensembl.org/index.html Design...polymerase II for transcription) are more effective for genome-editing events. In the light of these results, ...now, the tool is available for human and mouse genomes only and deals with C-terminus epitope [email protected]. Keep pipetting and happy genome editing! Pooran Dewari is a postdoc in Steve Pollard...research interests include optimizing CRISPR/Cas9 genome-editing methods and applying them to study brain...JD. 2017. Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog PostPublishedMarch 28, 2018, 6:52 p.m....SpCas9’s 3’ NGG PAM occurs frequently in GC-rich genomes, but a PAM is not always available near the locus...there is an NGG PAM every 42 bases in the human genome. SpCas9 targeting should be easy, right? Yes and...TTTV PAM, improve targeting in AT-rich loci or genomes. Cas9s from other species, like S. aureus and N...more complex than NGG, resulting in lower overall genome frequency. Despite the progress we’ve made, there...al. performed GUIDE-seq analysis to characterize genome-wide off-targets, since increasing PAM flexibility... -
CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems
TypeBlog PostPublishedJuly 23, 2020, 1:20 p.m....evolving. Variants of Cas proteins can be used for genome editing, activating gene expression, repressing...CRISPR system, N. meningitidis can also be used for genome editing. In this study, the labs found that anti-CRISPR... Conclusion CRISPR is a great way to make those genome edits. But, just as important is a way to flip ...Peng X (2018) Anti-CRISPR-Based and CRISPR-Based Genome Editing of Sulfolobus islandicus Rod-Shaped Virus... genome editing... -
Quick Guide to Working with Drosophila Part 1: Getting Started with Flies
TypeBlog PostPublishedJuly 13, 2017, 2:30 p.m.... crosses. Fly genomes Compared to humans and mice, flies have a relatively simple genome. They have four...gene? Can you find an ortholog in the Drosophila genome? Continue reading and I’ll show you how Drosophila...are diploid except for their germ cells (sets of genomic material are contributed by mother and father)....live animals yet, Harvard’s DRSC/TRiP- Functional Genomics Resources website has in vitro reagents and tools... -
Your Top Requested Plasmid in 2016!
TypeBlog PostPublishedJan. 11, 2017, 8:47 p.m....2016 was an exciting year for genome engineering research. A variety of new tools came out including ...SpCas9 is still going strong as the basis for many genome editing experiments. So, without further ado, the...describing them here. References 1.Ran, F. Ann, et al. "Genome engineering using the CRISPR-Cas9 system." Nature... -
New Tools Enable CRISPRa for Neuroscience Applications
TypeBlog PostPublishedJuly 2, 2019, 12:51 p.m....transcriptional activators to induce gene transcription at genomic sites targeted by specific sgRNAs. Its RNA-guided...nature makes CRISPRa much simpler to implement in genome-wide screens over cDNA libraries. However, using... the p65-HSF1 transcriptional activator to each genome-bound dCas9 (see Figure 1a for the transgenic construct...Because CRISPRa can be targeted anywhere in the genome, it is relatively straightforward to use CRISPRa...occur. CD8+ cells were then isolated and their genomic DNA sequenced. Guide RNAs targeting several known...demonstrated by the Qi Lab, which developed a subgenome-scale CRISPRa screening approach in embryonic ... -
New CRISPR Tools: Cas7-11 and PASTE
TypeBlog PostPublishedFeb. 17, 2022, 2 p.m...., amplified the CRISPR locus from the bacterial genome, and inserted it into E. coli to express and then...worth of DNA with CRISPR-driven precision into a genome, can be considered the fourth generation of CRISPR...fusions, using Prime to lay a “beacon” into the genome that a fused integrase would then integrate into...10-20% integration of inserts up to 36 kb into a genome. This integration was observed in non-dividing ...S. Gootenberg, Omar O. Abudayyeh. Drag-and-drop genome insertion without DNA cleavage with CRISPR-directed... -
Droplet Digital PCR for AAV Quantitation
TypeBlog PostPublishedJan. 16, 2020, 2:18 p.m.... end up with a titer of 4 x 1012 genome copies/mL or 8 x 1012 genome copies/mL and both results would ...dynamic range of the ddPCR is between 1 and 100,000 genome copies (GC) per reaction. Since AAV titers tend...can be used to examine the integrity of the viral genome (Furuta-Hanawa et al., 2019). Tips and tricks for...self-complementary adeno-associated viral vector genome titers by droplet digital PCR." Human gene therapy... -
A new platform for fast functional characterization of potentially cancerous mutations
TypeBlog PostPublishedMay 7, 2019, 12:39 p.m....Cancer Genome Atlas (TCGA), a catalog of genetic mutations responsible for cancer from genomics data, ...by developing a moderate throughput functional genomic platform to annotate genetic perturbations that... -
Plasmids 101: Screens vs. Selections
TypeBlog PostPublishedNov. 10, 2022, 2:15 p.m....small molecules/drugs. With the advent of CRISPR, genome-wide knockout screens are possible with gRNA libraries...and then outcomes are measured. While the entire genome can be knocked out with CRISPR, the entirety of...knocking out the genes of interest (or the whole genome), they knock down gene expression temporarily. ...resources on the Addgene Blog CRISPR libraries Genome-wide screens with CRISPR Lentiviral CRISPR libraries... -
Hot Plasmids - February 2022
TypeBlog PostPublishedFeb. 22, 2022, 2:15 p.m....Engineered miniature CRISPR-Cas system for mammalian genome regulation and editing Anti-FLAG Frankenbodies...Cas12a-based screening method was used to create a genome-wide library capable of performing similarly to...Engineered miniature CRISPR-Cas system for mammalian genome regulation and editing by: Rachel Leeson The large...applications. Figure 4: Overview of CasMINI for genome engineering applications. Image from Xiaoshu, et... -
Viral Vectors 101: Viral Vector Elements
TypeBlog PostPublishedJuly 25, 2023, 1 p.m....packaging to infiltrate and integrate into the host genome. First, it needs to be virally enveloped which ... and permanently insert the cargo into the host genome. Lentiviral-specific components Lentiviruses have...While AAV often does not integrate into the host genome (more on this below), it’s still used to deliver...for viral replication and also aids in infrequent genome integration at the AAVS1 locus in humans. Cap is... -
Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers
TypeBlog PostPublishedJuly 28, 2015, 2:30 p.m....understand the function of the non-coding portion of the genome. Mendenhall and Richard Myers of HudsonAlpha (where...define all of the functional elements in the human genome. The pair has focused considerable attention ... gene regulatory proteins and the places in the genome where they can be found more quickly and efficiently...transcription factors and figure out where they bind to the genome. We’ve been doing that for years using this assay... -
CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt
TypeBlog PostPublishedApril 22, 2015, 2:06 p.m....the ultimate objective of genome design.” It’s genetic engineering on a genomic scale, with the expectation...everything to the scale of building synthetic genomes or genome-scale construction projects. Not from the ... -
28 Hot Plasmid Technologies from 2015
TypeBlog PostPublishedDec. 23, 2015, 3:30 p.m....from a random genomic location than to express the tagged protein from a defined genomic location or from...FRB-NES) resulting in a rapamycin-inducible Cas9 for genome editing. Without rapamycin treatment, the Cas9(...greater temporal control over CRISPR/Cas9 mediated genome modification and gene expression. The plasmids ...has been targeted to a very specific site on the genome via the CRISPR system. The ease and flexibility...reporter gene, or (4) multiple sgRNAs with Cas9 for genome engineering. The versatility and ease of generating... Multicolor CRISPR labeling chromosomal loci in genome Thoru Pederson’s lab at the University of Massachusetts...tool enabling the imaging of multiple endogenous genomic loci simultaneously in live cells. This system ... -
Easi-CRISPR: Generating Knock-In and Conditional Mouse Models
TypeBlog PostPublishedApril 5, 2018, 12:42 p.m....method for in situ germline genome engineering using CRISPR nucleases. Genome Biol. 19 (2018) (1):25. DOI...CRISPR genome editing has made it easier to create knockout alleles in a variety of species, including...developed i-GONAD as a simple method for delivery of genome editing components to the germline. In contrast... and conditional alleles by CRISPR/Cas-mediated genome engineering." Cell 154 (2013):1370-9. DOI: 10.1016...long ssDNA donors and CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186/s13059-017... -
RaPID Detection of RNA-protein Interactions
TypeBlog PostPublishedAug. 16, 2018, 12:40 p.m....plasmids for labeling genomic regions Find CRISPR plasmids for purifying specific genomic regions ...the literature and analyzing data from The Cancer Genome Atlas, Ramanathan et al. found evidence that suggests...experimental identification of RNA-protein interactions. Genome Biology. PubMed PMID: 24467948 PubMed Central PMCID... -
X-CHIME: Context Dependent Germline Knockout in Immune Cells
TypeBlog PostPublishedJan. 16, 2024, 6:13 p.m....CRISPR101: Validating Your Genome Edit How to Design Your gRNA for CRISPR Genome Editing CRISPR Cheat Sheat...from Addgene.org CRISPR Guide CRISPR: Protocol for Genome Deletions in Mammalian Cell Lines ... -
Mapping the 4D nucleome with CRISPR/Cas9
TypeBlog PostPublishedAug. 11, 2015, 2:30 p.m....uses fluorescently labeled Cas9 to label multiple genomic loci in live cells. While other systems can be ...is a great step forward in live cell imaging of genomic loci. Ma et al. anticipate that the technique will...this publication at Addgene. Dynamic imaging of genomic loci in living human cells by an optimized CRISPR...