Multiplex Genome Engineering Using CRISPR/Cas Systems¶
Why this mattered¶
TBD
Abstract¶
Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. Lastly, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.
Related¶
- cite → RNA-Guided Human Genome Engineering via Cas9 — The multiplex CRISPR paper cites RNA-guided human genome engineering via Cas9 as evidence that Cas9 can be programmed for targeted editing in mammalian cells.
- cite → A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity — The multiplex CRISPR paper relies on the dual-RNA-guided Cas9 endonuclease mechanism as the programmable cutting system adapted for genome engineering.
- cite ← Genome engineering using the CRISPR-Cas9 system — Both papers develop CRISPR-Cas9 for multiplex genome engineering at multiple genomic loci.
- cite ← RNA-Guided Human Genome Engineering via Cas9 — RNA-guided human genome engineering is linked to multiplex CRISPR/Cas genome engineering through programmable Cas9 targeting of multiple genomic loci.