We narrowed to 1,018 results for: PLE;
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TypeBlog Post...loxP, black arrowheads) and one mutated pair (for example- lox511, white arrowheads). It is necessary to ...express GeneR only in the presence of Cre. In this example, Cre would first invert the reporter and GeneR ...2008. A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit ...
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Illuminating Epigenetics with A FRET Based Biosensor
TypeBlog Post...modification is chromatin remodeling. Chromatin is a complex of DNA and histone proteins that compacts cellular...different disease phenotypes, are two fascinating examples of epigenetics in the form of imprinting. In imprinting...have huge impacts in the way we study and treat complex or multifactorial diseases, enabling faster diagnosis... -
Cloning Mammalian Cells with the Agarose Method
TypeBlog Post...with knockouts of specific genes. Many years ago, people in the laboratory became frustrated with using ...procedure (illustrated in the YouTube video below) is simple: After obtaining antibiotic-resistant colonies,...initially, or using secondary biological screens. For example, for cancer researchers, if growth properties in... -
Quick Guide to Working with Drosophila Part 1: Getting Started with Flies
TypeBlog Post...Compared to humans and mice, flies have a relatively simple genome. They have four chromosomes: the X (and ...chromosomes essentially are messed up chromosomes with multiple inversions. They are unable to participate in ... dominant phenotypes visible on the flies. For example, second chromosome balancers often have the curly... -
PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs
TypeBlog Post...RNA (pegRNA). The pegRNA consists of a target-complementary spacer, a scaffold sequence, and an extension...screens with oligo libraries where pegRNAs are coupled to their target allowing us to edit and evaluate...efficiencies for diverse prime editing systems in multiple cell types. Cell, 186, 1–17. https://doi.org/10.1016... -
Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells
TypeBlog Post...AdV-mediated transduction of gRNA:Cas9 ribonucleoprotein complexes into transformed and non-transformed cells yielded...a potential platform that could be realized via simple switching of the relevant gRNAs and donor DNA Adenoviral...vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array... -
PITChing MMEJ as an Alternative Route for Gene Editing
TypeBlog Post.... For more mechanistic details on HDR and NHEJ, please see the linked blog posts. Compared to its counterparts...strategy looks very similar to HDR, but it is completed using much shorter sequence homologies. Abbreviated...Genome-Editing Based Approach for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: ... -
Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments
TypeBlog Post.... Here is an example of a plasmid with a downloadable protocol listed under “Supplemental Document”: gRNA_Cloning...of checking off each step as you painstakingly complete your experiment – one small victory in a long ... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog Post...ribonucleoprotein (RNP), consisting of the Cas9 protein in complex with a targeting gRNA, to your cells of interest...CRISPR experiment is the generation of the RNP complex. While one option is to purchase the Cas9 protein...bread wheat using CRISPR/Cas9 ribonucleoprotein complexes." Nature communications 8 (2017): 14261. PubMed... -
Fluorescent Proteins 101: Green Fluorescent Protein (GFP)
TypeBlog Post.... Now GFP and its fluorescent derivatives are a staple in the lab. GFP is used in research across a vast...importance of the entire GFP protein, which is almost completely required to maintain fluorescent activity; very...exist with only slight variations separating them. Please note that many fluorescent proteins found on the... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog Post...light inducible ion channels (plasmid # 20298 for example) have revolutionized Neuroscience. They are the...researcher exquisite control over expression of multiple genes in specific cells at specific times. Many.... “A FLEX Switch Targets Channelrhodopsin-2 to Multiple Cell Types for Imaging and Long-Range Circuit ... -
Antibodies 101: Reading a Flow Plot
TypeBlog Post...granularity, a proxy for nucleic complexity of the cell. The sample will have been labeled using a panel...parallel. Let’s imagine we’re looking for cells in a sample that are negative for marker A and positive for... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog Post...of DNA and both identified lymphocyte antigen 6 complex (LY6A) as a receptor for PHP.B. Read on to learn... the majority of astrocytes and neurons across multiple CNS regions. PHP.eB, an enhanced version of PHP.B... interested in using AAV-PHP.B vectors? See Supplementary Table 3 in Huang et al. for a list of PHP.B ... -
CRISPR-mediated Plant Base Editors
TypeBlog Post...polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers which enables us to target more than...Minkenberg, and Yinong Yang. "Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing...PMC5972399. 12. Molla, K., Shih, J. & Yang, Y. Multiplex CRISPR-mediated base editing in rice. In Genome... -
Adeno-associated Viruses (AAVs) for Genome Editing
TypeBlog Post...system frequently over the past fifteen years. Some examples from their lab include using AAV to introduce ... alleles of a gene, or which require modifying multiple genes simultaneously in the same cell, are more...by using a targeted nuclease. Asuri et al, for example, used zinc fingers to increase the efficiency of... -
3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells
TypeBlog Post...pluripotent stem cells (iPSCs). The CRISPR/Cas9 complex is very efficient at introducing double stranded...the PAM as possible (see below to find out why). Please check our recent studies for details (Paquet, Kwart... systems. References 1. Cong, Le, et al. "Multiplex genome engineering using CRISPR/Cas systems." ... -
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog Post...GCN4 binding scFv that’s fused to GFP. When the multiple copies of the scFv-GFP fusion bind the SunTag ..., Alitheen NBM, Hamid M (2012) scFv Antibody: Principles and Clinical Application. Clinical and Developmental....0000000000000145 Gąciarz A, Ruddock LW (2017) Complementarity determining regions and frameworks contribute... -
How to Design Your gRNA for CRISPR Genome Editing
TypeBlog Post... any gene of interest, one should require that multiple gRNAs of different sequences give rise to the ...but can often require difficult decisions. For example, would it be better to use a less-active gRNA that... in the interpretation of results by requiring multiple sequences targeting a gene to score in order to... -
CRISPR 101: Epigenetics and Editing the Epigenome
TypeBlog Post...allows for targeted delivery of Tet1 alone, or coupled with DNA oxidation and repair factors that allow...more frequently inherited by daughter cells. For example, KRAB induced silencing was transient and quickly...and context-dependent, it may make sense to try multiple CRISPR tools when setting up your experimental... -
Viral Vectors 101: AAV Serotypes and Tissue Tropism
TypeBlog Post...primary cellular receptor and sometimes one or multiple coreceptors, which may be present on different...and modifying capsid and genomic structures. For example, rAAV pseudotypes mix the genome of one AAV serotype... AAV8 AAV8 has high transduction efficiency in multiple tissues, rapid transgene expression, and can cross...