CRISPR gRNAs

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A partial CRISPR prime editing schematic cartoon. The parts of the prime editor and pegRNA are indicated. The prime editor consists of a Cas9n H840A nickase fused to a reverse transcriptase (RT), while the pegRNA includes spacer, scaffold, RT template, and primer binding site (PBS) sequences. The desired edit is part of the RT template. In the first step, these components form a complex, bind target DNA, and nick the Cas9 non-target strand. An arrow leads to additional steps not shown in this preview.
A cartoon showing a simplified Cas9 and gRNA binding to DNA. The 3’ end of the gRNA is the scaffold sequence, bound by Cas9, where some DNA substitution is tolerated. The 20 bases on the 5’ end of the gRNA are the spacer sequence, which hybridizes with the genomic DNA, base pairing to the target strand.   The non-target strand of genomic DNA includes the same sequence as the spacer. The PAM consists of the three bases immediately 3’ of this spacer-matching sequence.  Within the gRNA spacer, the 3’ half is the seed sequence, where no DNA substitution is tolerated. The 5’ half, also the 5’ end of the entire gRNA, is the tail sequence, where DNA substitution is tolerated.
screenshot of various PRIDICT webpages
Stabilizing gRNA Modifications
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
GoldenGate
Custom CRISPR Scren Schematic.png
Figure 1.png

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