CRISPR Human - Activation GRP 78

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

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4 years ago

4 years ago by Milena Alexeyeva Russian Federation

DNA insert using CRISPR

I would like to excise a large strand of DNA and insert a new one using CRISPR. My problem is that my strand will be a little over 1kb and I am not sure if this is going to be a limiting factor. Also, how long should the homology arms be for a region of this size?

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GRP 78 CRISPR Knockout and Activation + UltraCruz® Transfection Reagent

GRP 78 CRISPR Knockout and Activation

Santa Cruz Biotechnology

UltraCruz® Transfection Reagent

Santa Cruz Biotechnology

Upstream tips
If transfecting more than one plasmid (i.e., CRISPR/Cas9 KO Plasmid with HDR Plasmid), mix Plasmid DNA at equivalent ratios.
Protocol tips
Do not add antibiotics to the Plasmid Transfection Medium
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