CRISPR-CAS9 is a powerful technology for gene editing. It allows targeted modifications into the genome of the cell lines, IPSCs and ESCs. Despite the principle being pretty simple, in practice its use requires some expertise, and it can even turn into a time consuming adventure… That’s why the best way to benefit from it is most certainly through affordable services.
Many research labs would like to acquire and implement the CRISPR-CAS9 technology for their gene editing projects. Indeed, it’s a powerful system based on a simple principle: an endonuclease, the CAS9, driven onto a target site by a short guide RNA. There are so many strategies with benefits and drawbacks that is quite challenging to figure how to start out. Newcomers may be pushed into necessarily becoming experts before finding an efficient way to success. But what if you could use a complete and simple kit to facilitate your projects?
Capping is a required protection in 5′ of the mRNA that is necessary for biological activity and stability into the cells. There are already several methods of capping using mCap, ARCA or enzymatic methods. Unfortunately, they are limited in efficiency (such as mCap and ARCA), and can be costly (such as the enzymatic methods). Trilink Biotechnologies, the modified nucleic acid experts, have developped a new, far more efficient technology called CleanCap available in Europe via tebu-bio. [Read more…]
Vector-free CRISPR-CAS9 gene editing to accelerate therapeutic applications
A few years ago, Ayal Hendel et al (doi:10.1038/nbt.3290) published results revealing that chemical alterations to sgRNA enhance gene editing in primary cells. To demonstrate this, Matthew H Porteus’s team chose the targeted genes CCR5, HBB and IL2RG respectively involved in anti-HIV clinical trials, cell anaemia and thalassemia, and severe combined immunodeficiency. More recently, the same team tested several modified CAS9 mRNA. You can find the practical results on this poster introduced at the ASGC. [Read more…]
Shen et al (Nature Methods, 2017) explore and identify synthetic interactions among 73 cancer-associated genes. To perform their loss of function screen they combined CAS9-expressing cell lines with a sgRNA library of high titer lentiviral particles. Most of these gene interactions were subsequently validated by drug treatment. [Read more…]
Since the discovery of reprogramming factors in 2006 and the boom of CRISPR gene editing strategies, induced pluripotent stem cells (iPSC) have emerged as new cellular models. The development of 3D cell culture technologies has also contributed to the generation of induced Pluripotent Stem Cell (iPSC) derived cells, with unique applications from patient-specific drug responses testing, to regenerative medicine. I would like to introduce in this post a selection of reagents in this domain, a combination of both routine and innovative quality reagents, that I consider as bringing something extra to your stem cell research projects.
Today, I’d like to invite you to take a look at a highly efficient and useful kit, which brings together all the required components you need in a complete system for culturing and transfecting human pluripotent stem cells for gene editing.
The PluriQ™ G9™ Gene Editing System includes the G9™ Maintenance Medium and G9™ VTN Recombinant (vitronectin) plate coating for culturing human induced pluripotent (iPS) or embryonic stem (hES) cells in a manner that maximizes transfection by the included EditPro™ Stem Transfection Reagent to transfect genome editing constructs. [Read more…]
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas (CRISPR-associated) and Transcription Activator-Like Effector Nuclease (TALEN) are endonuclease based technologies aimed at developing targeted genome editing technologies.
CRISPR and TALEN provide Scientists with unique discovery tools for pathophysiology or genotype-phenotype studies by creating cellular models with gene knock-out, knock-in or tagging, promoter swapping, nucleotide substitution, protein truncation, reading frame disruption, modification of regulation by miRNA, genetic defect corrections…But, which one is the best for your application?