Joung Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids
RNA-Guided Nucleases ( RGNs ), based on naturally occurring Type II CRISPR-Cas systems, are programmable endonucleases that can be used to perform targeted genome editing. RGNs consist of two components: a short ~100 nt guide RNA ( gRNA ) that uses 20 variable nts at its 5’ end to base pair with a target genomic DNA sequence and a nuclease (e.g.—the Cas9 endonuclease) that cleaves the target DNA.
The Joung lab recently described gRNA and Cas9 expression vectors and validated the use of RNAs produced by these vectors for efficiently modifying endogenous genes in zebrafish (Hwang and Fu et al., Nat Biotechnol. 2013).
The Joung lab has also modified their web-based ZiFiT Targeter software to enable identification of potential RGN target sites and to facilitate design of short oligonucleotides needed to construct gRNA expression vectors. More information about RGNs and CRISPR-Cas systems can also be found at http://www.crispr-cas.org . Users are also encouraged to join the Genome Engineering newsgroup here .
Links to additional pages describing Joung Lab CRISPR-Cas/RGN reagents:
- Cpf1 expression plasmids (AsCpf1 and LbCpf1) for use in mammalian cells
- SpCas9 variants with no detectable off-target effects
- Modified PAM specifity of Staphylococcus aureus Cas9 (SaCas9 KKH variant)
- Engineered CRISPR-Cas9 nucleases with altered PAM specificities
- Dimeric CRISPR RNA-guided FokI Nucleases (RFNs)
- CRISPR-Cas/RGN expression plasmids for use in human cells
- CRISPR-Cas/RGN expression plasmids for use in zebrafish
- Guide RNA expression plasmids for the EGFP Reporter Gene
- Guide RNA expression plasmids for endogenous human genes
- Guide RNA expression plasmids for endogenous zebrafish genes