Nine months since the British vote to exit the European Union (“Brexit”), the UK science community’s initial dismay has given way to hard-boiled determination to limit the damage it will do to universities and research. On 29 March, Prime Minister Theresa May is expected to give formal notification of the UK’s intention to withdraw under Article 50 of the Lisbon Treaty, the constitutional basis of the EU. This will set in motion a 2-year period of intense negotiation on the terms of the UK’s divorce, and any future agreements with the EU—with research just one line item on a long list of issues to be resolved.
Most investigators get their engineered mice from colleagues or by purchasing them from commercial outfits like JAX or academic-based repositories. Popular engineered mice, such as JAX’s immunodeficient NOD scid gamma strains, sell for as little as a few hundred dollars, but a custommade mutant could cost as much as $20,000. By making the engineering of mice far simpler and cheaper, CRISPR opens the way for more labs to do it themselves. “When you made knockout mice before, you needed some skills,” says Rudolf Jaenisch at the Massachusetts Institute of Technology (MIT) in Cambridge. “Now, you don’t need them anymore. Any idiot can do it.
Bacterial adaptive immunity uses CRISPR (clustered regularly interspaced short palindromic repeats)–associated (Cas) proteins together with CRISPR transcripts for foreign DNA degradation. In type II CRISPR-Cas systems, activation of Cas9 endonuclease for DNA recognition upon guide RNA binding occurs by an unknown mechanism. Crystal structures of Cas9 bound to single-guide RNA reveal a conformation distinct from both the apo and DNA-bound states, in which the 10-nucleotide RNA “seed” sequence required for initial DNA interrogation is preordered in an A-form conformation. This segment of the guide RNA is essential for Cas9 to form a DNA recognition–competent structure that is poised to engage double-stranded DNA target sequences. We construe this as convergent evolution of a “seed” mechanism reminiscent of that used by Argonaute proteins during RNA interference in eukaryotes.
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