An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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 Schematic of targeted gene knock-out (left) and targeted DNA knock-in (right) by CRISPR/Cas9. Adenoviral CRISPR vectors (left) or CRISPR vectors and donor DNA vectors (right) are transduced to target cells. Within target cells, Cas9 machinery is transiently expressed and binds to target DNA. In the final step of targeted gene knock-out (left), a fragment is deleted from two homologous strands of DNA through error-prone non-homologous end joining (NHEJ). In targeted DNA knock-in, donor DNA is copied into the cut site through error-free homologous recombination (HR).
Multiplexed insertion is possible using a single plasmid that contains a multi-spacer CRISPR array. This encodes separate gRNAs that target different locations in the genome.
comic of illustrating Cas protein binding a gRNA and target DNA. The Cas protein is labelled CasM(onster) and looks like cookie monster. It has a quote bubble that says
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