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.

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.

Sequence engineering and chemical modification of CAS9 mRNA

Extract from poster “Maximizing Translation of Cas9 mRNA Therapeutics by Sequence Engineering and Chemical Modification”.

For transgenesis and therapeutic perspectives, vector-free delivery of the CAS9 for gene editing is highly preferable. It avoids the risk of hitting the genome due to the vector integration. Directly using CAS9 mRNA is a top option.

Furthermore, as a new promising class of therapeutic biologics, messenger RNA are the subject of numerous studies.

This year, Matthew Porteus’s and Trilink’s teams revealed key and practical results at the ASGC with a poster you can download below.

Notably, there is a large screen of the mRNA CAS9 modifications based on the indel formation. This interesting comparison includes also CAS9 RNP and reveals the high performance of the CleanCap U-depleted 5-moU CAS9 mRNA. We can also note that for the CleanCap CAS9 mRNA HPLC purification is not required.

“Maximizing Translation of Cas9 mRNA Therapeutics by Sequence Engineering and Chemical Modification”
Download your copy of the poster here.

Messenger RNAs – a promising class of new therapeutic biologics

As messenger RNAs (mRNAs) are easier to deliver into cells than plasmids or viral vectors, they are useful for non-dividing cells. Inversely to the vectors, they ensure genome integrity that is recommended for cell therapeutics. mRNA are also well adapted to transient expression as required for cellular reprogramming, gene editing, and vaccines.

You might like to take a look at this poster, presented by Trilink (renowned modified nucleic acid experts) at the Keystone symposia last March (Pattern Recognition Signaling: From Innate Immunity to Inflammatory Disease). They share results on how to optimize messenger RNA for therapeutic activity.

“Innate Immune focused approaches to maximize messenger RNA therapeutic activity”
Download your copy of the poster here.