Tuesday, 17 February 2015

Genome Editing Stabilised mRNA

Genome Editing mRNAs

Plasmids and viral vectors are traditionally used in CRISPR genome editing to express the required proteins. However, there is a risk in using DNA instead of mRNA: Double stranded DNA breaks catalyse insertion of DNA at the cut site. At some substantial frequency, the protein expression vectors can integrate which can lead to continuous expression of the nuclease or a previously silent sequence.

Now mRNA is becoming a powerful tool in genome editing. With no risk of insertional mutagenesis, mRNA can be used to transiently express the required proteins. Wild-type Cas9 mRNA is stabilised non-immunogenic messenger RNA (mRNA) that has been designed to induce a high level of expression of wild-type Cas9 protein. This creates a double stranded break at a target site delineated by RNA guide sequences.

This mRNA is produced by in vitro transcription, stabilised at the 5’ end by modified nucleotides capping and contain a poly(A) tail at the 3’ end. Cas9 mRNA has been optimised for improved stability, improved performance, and to lower the cellular innate immunity response.

mRNA transfection provides several advantages:

-No need for nuclear uptake - protein expression occurs directly in cytoplasm
-Faster protein expression than DNA transfection
-No genomic integration
-Perfect for transfecting slow or non-dividing cells
-Protein expression in a total promoter-independent manner
-Transient transfection: mRNA based expression of proteins is sustained for a limited time.

Product Code
Description
Size
Cas9 mRNA
20µg
Cas9 mRNA
100µg
CRE mRNA
20µg
CRE mRNA
100µg


Caltag Medsystems are the UK distributor for OZ Bioscience. For further information about their products, please visit our website, www.caltagmedsystems.co.uk, call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk.

1 comment:

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