Skip to main content

Genome Engineering Services

VGEF Puts CRISPR Gene Editing at Your Fingertips

VGEF is your one-stop shop for CRISPR gene editing services. We offer custom services in immortalised cells, primary human/murine cells and iPSC in support of fundamental and preclinical biomedical research.

Building on exciting recent developments in the field of gene editing and genome engineering, we are pleased to offer a wide range of services based on the state of the art CRISPR technology, and more! We can confidently tackle a wide range of applications, from the ‘standard’ CRISPR services - such as gene knock-out, knock-in, and the introduction of point mutations - to gene expression modulation, epigenome and RNA editing.

Our expertise in CRISPR gene editing is combined with access to the latest transfection techniques. These include Amaxa 4D electroporation, and viral vector transduction techniques which allow us to achieve superior gene editing efficiency in a variety of cells, at lower cost and in shorter time. In other words, we do everything to make YOUR research faster, cheaper and more competitive.

Not sure where to begin? Not to worry! We offer consultation and process optimization services to help you tackle even the most challenging of gene editing projects.

Gene Editing technologies currently available at VGEF

While the Streptococcus pyogenes version of Cas9 (spCas9) was the first CRISPR-related nuclease to be used for gene editing, and remains the most widely used to this day, many exciting new enzymes have since been discovered and developed into research tools. These include the Staphylococcus aureus version of Cas9 (saCas9) and the Cas12 class of nucleases (previously known as Cpf1) which adds new options to the current gene-editing toolbox, such as targeting a T-rich PAM and the introduction of sticky-ended dsDNA breaks.

These enzymes are the product of great ingenuity and a tremendous amount of hard work, and new developments are emerging at a staggering pace. 

To help navigate the current gene editing landscape we have compiled a brief bibliography summarizing some of the key CRISPR technologies.

More information on CRISPR Nucleueases is available here.

CRISPR Knock-out

One of the most frequent applications of CRISPR is gene knock-out, which utilises CRISPR Cas9 machinery to introduce targeted dsDNA breaks and relies on the non-homologous end joining (NHEJ) pathway to re-join dsDNA ends, resulting in random insertions or deletions known collectively as INDELS. These in turn lead to a frameshift mutation resulting in a loss of function of the targeted gene.

VGEF has access to a comprehensive collection of nuclease variants and has hands-on experience with a wide range of gene-editing strategies. VGEF strives to ensure the highest rate of success for the fastest turnaround and lowest cost. We achieve this through an ongoing dialogue with our customers to ensure we understand your experimental needs and select the best strategy for you. The table below lists some examples of the most common strategies we employ.

Don’t see what you need? Not to worry! Simply contact us for a free consultation to find the approach that’s right for you.

CRISPR Knock-in

‘Knock-in’ is an umbrella term which encompasses a variety of CRISPR applications, all of which involve the introduction of a particular piece of foreign DNA into the genome of a living cell. This could be anything from a point mutation, a protein domain, a reporter tag, exogenous gene or even an entire expression cassette (complete with its own promoter and polyA signal). This is achieved by providing a repair template along with the usual CRISPR Cas9 machinery, which carries the exogenous DNA sequence flanked by sequences that are homologous to the region surrounding the dsDNA break.

Unlike CRISPR knock-out, knock-in projects tend to be a lot more complex and have a considerably lower rate of success. This is driven in part by the necessity of carrying out DNA cleavage as close to the desired point of insertion as possible, greatly increasing the difficulty of selecting optimal gRNAs. However, our large collection of nucleases gives us access to a variety of PAM sequences, allowing us much greater freedom to carry out targeted insertions. Browse the table below for a few strategies we use at VGEF for CRISPR Knock-In.

Don’t see what you need? Not to worry! Simply contact us for a free consultation to find the approach that’s right for you.

Beyond CRISPR gene editing

One of the most exciting technologies to emerge from the CRISPR revolution has been the development of dCas9-related tools and applications. Scientists realized they could inactivate the cleavage activity of Cas9 (known as ‘dead Cas9’, or ‘dCas9) without affecting its ability to specifically recognise DNA fragments in the same fully-customised and targeted fashion. Thus, by attaching various effector domains to dCas9, we are now able to act upon genomic DNA in a variety of ways, such as targeting promoters to boost or lower endogenous gene expression, methylate or demethylate DNA or histones to modify the epigenome, or use fluorescently label dCas9 to visualise particular DNA sequences.

By taking advantage of the dCas9 SunTag system (Tannenbaum et al, 2014) VGEF is ready to tackle a variety of dCas9-related applications. Have an experiment in mind? Get in touch with us and find out what tools we have available to help YOU achieve YOUR unique goals.

Don’t see what you like? Not to worry! Simply contact us for a free consultation to find the approach that’s right for you.