Top 5 2017 Best Labs for Your CRISPR Cas9 Analysis

CRISPR (pronounced “crisper”), stands for Clustered Regularly Interspaced Short Palindromic Repeat. Short repeated DNA sequences in the genomes of bacteria and other microorganisms are organized in clusters. This explains the name.

CRISPR was inspired by bacteria’s defense mechanisms from viral attacks.The principle is simple. Bacteria take “bits” of the invading virus’ DNA. They splice it in short sequences with an enzyme called Cas. The newly-formed short sequences are called CRISPR. RNA copies of these sequences are made by the bacteria. Those sequences will help recognize the virus’ DNA and prevent future invasions.

Targeted, genetic modification in humans is no longer science fiction. In 2012, scientists turned CRISPR from a bacterial shield into a gene-editing tool. By “feeding” Cas9 the right sequence or modified guide RNA, scientists can cut and paste parts of a DNA sequence anywhere into the targeted genome.

This technique gives genetic biologists a powerful tool for gene editing. Scientists can change or add DNA within the cell.

You can save time and efforts by having genome engineering experts subcontractors develop your CRISPR/Cas9.

Here is a review of the subcontractors you can contact if you are interested in outsourcing your CRISPR Cas9 analysis.

Creative Bioarray

• From the USA •

Found in 2004, Creative Bioarray is an innovative biotechnology company.

They provide a wide range of pieces of equipment.

They also provide products such as cell culture medium and reagents, normal human and animal cells, probes for FISH experiment, tissue arrays, and microorganisms.

And they provide services such as biosample services, histology services, and genome manipulation by correcting mutations in disease-model iPSCs (pluripotent stem cells) using CRISPR/ Cas9.

Cyagen Biosciences

• From the USA •

Founded in 2006, Cyagen Biosciences is a contract research organization and cell culture product manufacturer. Based in the USA, with offices in Germany and China.

They specialize in molecular biology tools and manufacturer of research-use stem cells and related culture reagents. The company provides custom-designed transgenic and knockout mouse services.

They offer:

• BAC and PiggyBac transgenic services.
• CRISPR-mediated mouse and rat genome editing.
• CRISPR-based, large-fragment knockin, at any locus, and conditional knockout mice generation.
• Turboknockout® - an ES cell-mediated service that provides you with a conditional knockout.
• Reporter knockin and humanization mouse models.
• Vectorbuilder - a new online platform to design and order your custom vectors.

Cyagen specializes in mutation mouse model generation via CRISPR/Cas9-mediated genome editing technology. They use co-injection of nuclease and oligo donors carrying the desired mutation and homology arms.

StemCore

• Affiliated to the__University of Queensland • From Australia

Stem Cells is based at the Australian Institute for Bioengineering and Nanotechnology at The University of Queensland.

It is a not-for-profit biotechnology entity in the field of regenerative medicine.

They offer services, comprehensive training workshops and support of annual Cell Reprogramming Australia conferences.

This company enables stem cell research by providing comprehensive training workshops, and also foster networking among stem cell scientists to boost the profile of Australian regenerative medicine.

Stem Cells offers a proprietary bank of human embryonic and induced pluripotent stem cell lines.

Their services include:

• iPSC generation and characterization.
• Genome-editing using CRISPR-Cas9.
• Stem cell tri-lineage differentiation.
• Mouse embryonic fibroblasts (MEFs).
• DNA Karyotyping.
• SNP Genotyping.
• Mycoplasma Testing.
• Teratoma Assay.

Diagenode

• From Belgium •

Diagenode is a global provider for epigenetics research, biological sample preparation, and diagnostics assays based in Liege, Belgium, and New Jersey, USA.

This company offers innovative Bioruptor® shearing and IP-Star® automation instruments, reagent kits, and high-quality antibodies to streamline DNA methylation, ChIP, and ChIP-seq workflows.

At Diagenode, the latest innovations is a full automation system, the Megaruptor shearing system for long fragment generation in sequencing, and epigenetics assay services.

They can perform different CRISPR/Cas analysis:

• Checking the transfection success of the Cas9 protein, by checking Cas9 expression level using Western blot with validated anti-Cas9 antibodies.
• Checking that the Cas9 protein was delivered to the nucleus, by determining the nuclear localization of Cas9 protein via IF or IHC using validated anti-Cas9 antibodies.
• Checking the Cas9 transient and stable expression level, by using Western blot and anti-Cas9 antibodies.
• Checking the binding specificity of Cas9, with an sgRNA of choice by using the ChIP-grade anti-Cas9 antibody and the primers for targeted and non-targeted region to see if Cas9 is bound to the correct region.

Microsynth

• From Switzerland •

Microsynth is a Swiss company, specialized in services related to DNA/RNA synthesis, various kinds of DNA/RNA analyzes and molecular biology project outsourcing.

Microsynth offers a broad range of analysis for CRISPR/Cas9 application:

• Synthesis of tracrRNA and crRNA oligonucleotides.
• Synthesis of DNA oligonucleotides for molecular cloning of the target-specific region of your gRNA.
• Sanger sequencing: Sequence verification of your CRISPR plasmid.
• Analysis of specific CRISPR/CAS9 edited clones by PCR, cloning and Sanger sequencing.
• Next-generation sequencing: Validation of single CRISPR/Cas9 targets, using an amplicon deep-sequencing approach.
• Measurement of expression levels of targeted genes using RTPCR.
• Detection of the edits in CRISPR/Cas9 screenings.

Microsynth has also developed protocols for DNA isolation, PCR and sequencing for CRISPR/Cas9 screening.