Articles, announcements and insights from Horizon Discovery

How to bank cell lines - Our recommended protocol

Jun 10, 2016 2:26:45 PM No Comments

Whether it's a stock you've expanded following it's arrival from the cell bank, or a clone you've carefully nurtured from the single cell, banking down your cells in the right way is crucial if you're going to be able to return to them time and again, and revive them quickly so that you can get on with your experiments.

Here's our protocol for banking human cell lines:

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[Protocol] Optimizing transfection for genome editing

Jun 10, 2016 2:25:54 PM 1 Comment

Transfection or electroporation is used to efficiently introduce the nucleic acids required for CRISPR cell line engineering into a cell line. The type and number of nucleic acids (usually plasmids) being introduced into a cell line will depend on the engineering event being undertaken. At its most simple, gene knockouts can be achieved by transfection of a plasmid expressing wild-type Cas9 along with a guide RNA (gRNA) to the gene that is being knocked out.

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[Protocol] Harvesting and analyzing clones following CRISPR gene editing

Jun 10, 2016 2:25:18 PM No Comments Comment

If you've followed our guide to planning a successful genome editing experiment, then you'll hopefully be working in optimal conditions, and have a good idea of your guide's editing efficiency. This number should give you a rough idea of how many clones you're going to have to screen to find a targeted clones. Keep in mind the following:

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[Protocol] Expansion of cell line clones from single cells

Jun 10, 2016 2:23:34 PM No Comments Comment

Once a clone has been identified positive for targeting in the initial screen, the cells should be expanded to a 48-well plate. Expansion of the cells can usually occur at 1-4 days post screen.

Read More Cell lines, Gene editing, Protocols

Rapid identification of potential drug targets using endogenous reporter cell lines and siRNA screening

Jun 10, 2016 2:22:13 PM No Comments Comment

We have combined our proprietary genome editing technology with large scale RNAi screening capabilities to allow identification of novel drug targets.

Read More Cell lines, siRNA Screening, Endogenous reporters

Developing Cellular Reporter Technologies Using Horizon's Cell Models

Jun 10, 2016 2:20:39 PM No Comments Comment

Isogenic cell lines provide genetically defined, patient-relevant, predictive in vitro models of genetic disease. We are further extending their application within targeted drug discovery with the development of new reporter disease models using our gene engineering technology. These models combine Horizon's cell lines with the endogenous gene reporting capabilities in the form of NanoLuc® luciferase and HaloTag® reporter technologies.

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Using endogenous reporter cell lines in high-throughput screening

Jun 10, 2016 2:09:44 PM No Comments Comment

Reporter gene assays are widely used to study the regulation of gene expression. We have developed a suite of endogenous reporter cell lines which measure natural levels of protein expression and promoter activity. By measuring at the endogenous level, this system provides an advantage over other technologies which use exogenous plasmid-based overexpression systems.

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Improving Gene Editing Success with An Optimised Cell Line Screening Procedure

Jun 10, 2016 2:06:25 PM No Comments Comment

There exist now a range of techniques to perform genome editing, such as ZFN, CRISPR, TALENS and AAV, each with their own strengths and weaknesses. However, one consistent element that has a significant impact on the success of that editing event when generating an isogenic cell line is the choice of parental cell line to be engineered.

Read More Cell lines, Gene editing

Enhancing the efficiency of knockins by DNA mismatch repair suppression and negative selection

Jun 10, 2016 2:04:22 PM No Comments Comment

Recombinant adeno associated virus (rAAV) is a precise and effective method to introduce defined changes into endogenous genes and rAAV vectors can stimulate homologous recombination (HR) up to 1000-fold over that seen using plasmids.

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Improved gene editing efficiencies using AAV and CRISPR combined

Jun 10, 2016 2:02:04 PM No Comments Comment

Nuclease based approaches like CRISPR-Cas9, ZFNs and TALENs facilitate targeted modification of genomes by inducing double-strand breaks (DSBs) within chromosomes at specified locations. This stimulates the natural DNA-repair mechanisms of homologous recombination and non-homologous end joining.

Plasmid DNA, PCR products, and single stranded oligonucleotides are routinely used as donors to introduce specific changes at the DSB site. The efficiency of introducing a desired change is dependent on many factors including

  • the type of donor
  • the length of homology
  • the complexity of the desired change
  • characteristics specific to the cell line

We have looked at whether rAAV vectors can be used as donors for DNA modification to obtain higher efficiencies than seen with other donor approaches.

Read More Cell lines, Gene editing

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