CRISPR Full Service

benefits.

CRISPR Strategy Design

A key step of any CRISPR experiment. We know the potential pitfalls.

Optimized gRNA Selection

Use of tools providing better gRNA efficiency predictions.

Sequence Verification

Of gRNA target sites, homology arms and constructs. Avoids bad surprises.

the service.

CRISPR is a powerful genome editing technology that allows us to analyze genes and regulatory elements with unprecedented precision. Although elegant, CRISPR is not always easy to use.

To help, we offer a CRISPR Full Service that includes all steps from design to the final CRISPRed fly strain.

The challenges of implementing the CRISPR technology have become obvious from the many and diverse injection projects that we have been involved with. The CRISPR workflow involves multiple steps, and even small missteps in this workflow can result in weeks or months of wasted time and effort.

our approach.

Whereas most CRISPR experiments still use early technologies (e.g., several plasmids for gRNAs and donor constructs), we use advanced methods and apply specific tests.

single plasmid

Whenever appropriate all gRNAs and knockin/knockout components are contained in one physical DNA entity.

in vivo linearized templates

Use of in vivo linearized templates rather than supercoiled plasmids (linearization enhances HDR).

target site/homology arms

Target site and homology arm validation (i.e. PCR and sequencing).

selection of gRNAs

Selection of gRNAs with bioinformatic tools providing better gRNA efficiency predictions.

in vivo gRNA

in vivo gRNA testing (optional)

sequence-verified constructs

Use of sequence-verified constructs.

One key step is the CRISPR design, and it is the place where many CRISPR experiments struggle. A second key step is the selection of efficient gRNAs. We will use our expertise to design the best strategy for you.

how does it work.

First

Client provides key information

CRISPR goal, target (e.g. gene), special requirements

Second

We'll get in touch with you

Open questions, etc. We provide a quote

Third

CRISPR Design

gRNA selection, construct design

Fourth

PCR/sequence validation

gRNA site/homology arms

Fifth

Cloning

gRNA together with donor

Sixth

Injection & screening

(Optional)

PCR validation

Seventh

Analysis and reporting

The whole workflow takes roughly 15–18 weeks, depending on complexity and what services are included.

get a CRISPR full service quote.

Please describe the goal of your CRISPR experiment and your current editing ideas to achieve this goal in the message field below.

other services.

Explore a range of specialized solutions tailored to your genome editing needs.

CRISPR Injections

CRISPR is a powerful genome editing technology.

PhiC31

Injection of your construct into a phiC31-attP strain.

P-element

Injection with a helper plasmid and the P-element.

DNA

We will dilute the DNA to an injection concentration.

FAQs

What happens if you don’t get any CRISPR transformants with the CRISPR Full Service?

There are a number of parameters that are relevant for the success of a HDR-based CRISPR experiment. Some of them can be influenced and optimized (e.g. tools/reagents used, quality of the reagents, selection of gRNAs), whereas others can often not be influenced (e.g. the accessibility of the CRISPR target locus due to chromatin structure). Based on our and others experience and literature, HDR-based knock-ins in Drosophila succeed up to ~80%.

In other words, it is currently impossible to guarantee that every HDR-based CRISPR experiment will work. If we are in charge of the CRISPR project (CRISPR Full Service) we will invest considerable time and efforts into evaluating your project and find a feasible strategy to make the project work.

If our first approach does not work we will try a second time. Depending on the specific case we will either try a different design, use different tools/reagents or use alternative gRNAs, without any additional costs for this second attempt. If the second attempt fails we will discuss with you how to proceed.

Note, the free second attempt only applies for marker-based CRISPR projects (e.g. 3xP3-RFP) and not for PCR-based screenings.