The Twelve Days of CRISPR

By Jennifer Tsang

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Using CRISPR in C. elegans to knock-in FLAG tag.
A schematic of the cell membrane with a GPCR and the transducer complex. When the GPCR binds the ligand, the transducer complex dissociates resulting in a decrease in BRET.
 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.
The RepCap plasmid consists of three promoters within the rep coding region that encodes four different Rep proteins. This region also encodes three different VP proteins, the MAAP protein, and the AAP protein.
A day in the life of a software product manager infographic
Schematic of the human MECP2 locus and MECP2 protein.

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