110 GENOME EDITING
Turning to CRISPRs for genome editing
Genome editing has taken a leap forward as the benefits of clustered regularly interspaced short palindromic repeats (CRISPRs) technology become more widely known. Sean Ottewell reports.
Édition du génome a pris un bond en avant que les avantages de cluster répétitions palindromiques courts régulièrement espacées (CRISPRs) la technologie soit plus largement connue. Rapporte Sean Ottewell.
Genome Editing hat einen Sprung nach vorn, wie die Vorteile der Cluster regelmäßig beabstandeten kurzen palindromischen Repeats (CRISPRs) Technologie besser bekannt gemacht. Sean Ottewell berichtet.
interspaced short palindromic repeats)/Cas9 system for genome editing.
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Fig. 1. The GeneArt CRISPR product portfolio is designed to offer many options for complete flexibility and ease of use.
Tis is an RNA guided DNA endonuclease system that has two key components, the Cas9 protein that serves as an DNA endonuclease and a non-coding RNA component called guide RNA (gRNA) that confers target specificity. Te CRISPR/Cas9 system works in conjunction with endogenous repair mechanisms to alter DNA within a cell.
ene editing – the modification of an organism’s genome
via a specific, targeted approach – is among the fastest growing applications for molecular biologists. Now, new tool kits are available to researchers that enable them to quickly make specific changes to DNA in a variety of cells.
For example, GeneArt CRISPR products from Termo Fisher Scientific leverage the type II CRISPR (clustered regulatory
Te CRISPR product portfolio is designed to offer many options for complete flexibility and ease of use (Fig. 1). Tese include, for example, GeneArt CRISPR nuclease mRNA, which is ready to transfect wild type Cas9 in mRNA format. Te mRNA format is best for multiplexed screening of gRNA sequences, applications that require delivery by microinjection, and easier dosage optimisation of Cas9 protein to gRNA in the cell. Cas9 mRNA is co-transfected with one or more custom GeneArt CRISPR strings DNA encoding target-specific gRNA or in vitro transcribed gRNA which removes promoter constraints and is a better option for difficult to transfect cells such as induced pluripotent stem cells, or iPSCs (induced pluripotent stem cells).
A second option is the GeneArt CRISPR nuclease vector kits which offer reporter and enrichment tags for tracking gene editing experiments and are great for cloning and re-use of confirmed gRNA sequences. Te GeneArt CRISPR nuclease vector with CD4 enrichment is a vector system for expression of the functional components of the
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CRISPR/Cas9 technology with a CD4 reporter. Te CD4 reporter offers magnetic bead-based sorting/enrichment of Cas9 and CRISPR expressing cells using Dynabeads CD4 magnetic beads. Transfection efficiency can also be tracked using anti- CD4 fluorescent antibodies. Te GeneArt CRISPR nuclease vector with orange fluorescent protein (OFP) reporter is a similar expression vector system with an (OFP) reporter. Te OFP reporter allows for fluorescence-based tracking of transfection efficiency, as well as fluorescence activated cell sorting (FACS)-based sorting/ enrichment of Cas9 and CRISPR expressing cells.
GeneArt CRISPRs can be used with other gene editing tools, such as the GeneArt Genomic Detection Cleavage Kit, providing a complete solution for design of synthetic genes, circuits and detection. Termo Fisher can also design and clone CRISPRs for customers through the custom services team.
“CRISPRs provide a way to quickly and easily target those genomic sites a researcher wishes to modify in order to engineer cells,” said Helge Bastian, vice president for synthetic biology, life sciences solutions, Termo Fisher Scientific. “Our new CRISPR kit format, along with our transfection reagents, offers a simplified workflow that enables the less-experienced user to easily carry out gene editing.”
Meanwhile Sigma Aldrich has launched Sigma CRISPRs, its latest offering to the global research community. “As the first company to commercially offer targeted genome editing
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