Recycling has always been a smart idea, and nature has its own processes to ensure that waste is kept to a minimum. As Professor Ohsumi discovered, autophagy is the cells way of degrading and recycling cellular components, allowing it to adapt to nutritional deficiency or other environmental influences. Professor Yoshinori Ohsumi, honorary professor and leader of the Cell Biology Unit at the Tokyo Institute of Technology, has been studying autophagy for 27 years. This year's Nobel Laureate discovered and elucidated mechanisms underlying autophagy, according to the Press Release from The Nobel Assembly at Karolinska Institutet.
On Saturday 10th September 2016, a team of intrepid volunteers defied the rain and blustery weather to compete in the Cambridge Dragon Boat Festival 2016 in aid of ACT (Addenbrooke’s Charitable Trust). Proudly representing Horizon Discovery, in their branded t-shirts, were our 11 person team who, unlike several of the more experienced teams, gave an excellent account of themselves in the inclement conditions and managed to stay afloat throughout.
On 5th September at our offices in Cambridge, Lord-Lieutenant, Sir Hugh Duberly KCVO, CBE presented Dr Darrin Disley, CEO of Horizon Discovery, with a Queens Award for Enterprise Promotion.
Darrin was nominated for the Queen's Award for Enterprise Promotion by Linda Allan. Her Majesty The Queen approved the Prime Minister's recommendation that Darrin should receive the Award.
Which Cell line models to use?
Sourcing biological materials that will acurately represent disease biology can be a time consuming and frustrating proceedure. Here we discuss:
- the options available
- and the merits and disavantages of each
- how gene editing technology can provide bespoke cell models of disease
Increasingly the literature suggests that CRISPR's potential for off-target effects is not as “bad” as originally thought. Here are a few things to set your mind at ease if you’re working with the CRISPR-Cas9 system:
Early on in our CRISPR use at Horizon, we learned that that when it comes to guide RNA design not all sgRNAs are created equal. In fact what makes a good guide is still the topic of a great deal of research in the field – and aside of the consensus that a guide with a guanine at position -1 is more likely to have a higher activity, we are still a way from being 100% confident in predicting in silico guide activity in vitro/vivo.
The CRISPR/Cas9 system has been rapidly adapted to practically every model system for its ease to generate and high efficiencies to cleave target DNA. But unlike our experience with Zinc Finger Nucleases, in the human, rat and mouse cell lines we tried successful co-transfection of Cas9 mRNA and sgRNA was cell-line dependent, and often resulted in either very low or no cleavage activities.
However, sequential transfection of cells with Cas9 DNA first, and sgRNA followed 24 hrs later, reliably produced good level of activity, indicating the requirement of Cas9 presence at the time of introduction of sgRNA. Not surprisingly, creation of a cell line stably expressing Cas9 led to consistently high cleavage activities upon transfection of sgRNAs. Transfection of recombinant Cas9 protein pre-complexed with sgRNA (ribonucleoprotein particles, or RNPs) led to efficient cleavage as well.
On the other hand, when Cas9 mRNA and sgRNAs are co-microinjected into single cell embryos, it produces target cleavage as efficiently as RNPs to produce straight KOs and large deletions between two target sites, again raising a question of local concentrations of Cas9 protein and sgRNA.
Below we summarize some of the work we've done optimizing delivery of CRISPR-Cas9, which which can be read in full publication form in Human Gene Therapy here.
Here you'll find a complete list of our most frequently asked questions relating to X-MAN cell lines. How are they made, how are they validated, and how can you use them? Read on to find out.
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: