Manipulating iPSCs with CRISPR – Functional knockouts, fluorescent tagging, and expression activation

15 Oct 2020

This application note by Horizon Discovery presents the ability to modify the genome and modulate expression of genes within a human induced pluripotent stem (iPS) cell line with Dharmacon™ Edit-R CRISPR reagents.

CRISPR Screening Services

CRISPR Screening Services Horizon offers comprehensive CRISPR screening services from screen design and cell line selection to sophisticated bioinformatics analysis of the screen results. Our best-in-class CRISPR KO (knockout), CRISPRi (interference) and CRISPRa (activation) screening platforms can significantly advance our customers’ drug development programs by offering the highest quality and highest confidence in their screening results. Data analysis services, including sample preparation, Next-Gen Sequencing, and hit calling, can be provided for users of Edit-R Lentiviral sgRNA Pooled Screening Libraries. Simply provide whole cell pellets for each of your samples and we’ll do the rest. We can also help with the NGS data analysis and hit calling for data created using the Edit-R Pooled sgRNA Indexing PCR and Sequencing Primer Kits. Types of CRISPR Screen Expertise includes whole-genome and custom targeted drug resistance and sensitivity screens, synthetic lethality target discovery screens and complex phenotypic screens using a FACS-based readout. We have optimized 50+ cell lines for CRISPR screening and have completed over 300 screens to date. We offer a choice of CRISPR screen types depending on the expression effect you require: CRISPR KO screening with complete loss of gene expression provides the maximal window for phenotypic effect and high statistical power for hit discovery CRISPRi screening represses expression rather than completely knocking out the target gene and is ideally suited to study druggability and to evaluate the function of genes that when knocked-out are essential or that are amplified CRISPRa screening amplifies gene expression in its endogenous context and enables for the first time, to study activation-linked responses on a genome-wide level Dual CRISPRi & CRISPRa screening

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DNA / RNA Extraction and PurificationPurified DNA and RNA are required for numerous downstream molecular biology applications. Consequently, the importance of high-quality DNA/RNA extraction and purification equipment cannot be underestimated. Many purification kits are available and are typically optimized for nucleic acid type and source, including plasmid DNA, genomic DNA, mRNA, RNA and viral nucleic acid purification kits. Automated extraction and purification of nucleic acids can be implemented with magnetic bead separator instruments or high-throughput purification workstations. Find the best DNA/RNA extraction and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA SequencingDNA sequencing, such as sanger sequencing, is a biological technique that determines the precise order of nucleotide bases in a fragment or template of DNA. DNA sequencers and genetic analyzers are based on capillary electrophoresis, where labeled DNA fragments are electrophoretically separated by size as they migrate through a polymer. Find the best DNA sequencing products, including DNA sequencing kits, genomic libraries and genetic identity kits in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Gene Expression and Molecular CloningMolecular cloning is a set of techniques that utilizes vectors to transfer recombinant DNA into host cells and is an essential tool for investigating the expression of genes and proteins in bacterial or mammalian cells. A variety of vectors optimized for gene cloning and expression in a range of host organisms are available, alongside competent cells for genetic replication. Here, you can explore a range of molecular tools, high-quality genomic and cDNA libraries, premade clones, transformation and transfection reagents and mutagenesis or gene expression detection assays and expression arrays. Find the best gene expression and molecular cloning products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA / RNA QuantificationDetection and quantification of nucleic acids is important in molecular biology, cloning, expression, forensics and clinical diagnostics. Nucleic acids can be detected by labeling with colorimetric, fluorescent or radio labels and using in situ hybridization kits to identify specific sequences. Multiple nucleic acids can be detected and quantified at once using RNA / DNA detection beads or RNA / DNA microarrays. Find the best DNA / RNA Quantification products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNADeoxyribonucleic Acid (DNA) is the main component of chromosomes and the carrier of genetic information of living organisms. Find out here about PCR, NGS, ChIP-Seq, gel imaging, and many other techniques which can be used for the analysis of DNA.Gene EditingGene editing is the precise alteration to an organism's DNA, with CRISPR being the most well-known technique. It is used in a wide range of applications, including the development of disease models and gene therapy, and crop improvement. Browse our peer-reviewed product directory to find the best gene editing solutions, compare products, check reviews, and get pricing directly from manufacturers.CRISPRCRISPR technology enables precise editing of genes, allowing scientists to modify DNA at specific locations. This revolutionary tool is used in genetic research, drug development, and gene therapy. CRISPR has applications in agriculture, disease treatment, and creating genetically modified organisms (GMOs). Explore CRISPR solutions in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.
Manipulating iPSCs with CRISPR – Functional knockouts, fluorescent tagging, and expression activation