Incontro, Salvatore; Asensio, Cedric S; Edwards, Robert H; Nicoll, Roger A. Neuron; Cambridge Vol. 83, Iss. 5, (Sep 3, 2014): 1051-7.DOI:10.1016/j.neuron.2014.07.043

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Historically, pharmacology provided most of the initial insight into function of central synapses. Selective antagonists for neurotransmitter receptors, which have the advantage of rapid and reversible actions, established the role of AMPA and NMDA receptors in synaptic transmission and plasticity (Collingridge et al., 1983; Hestrin et al., 1990; Watkins et al., 1990 ). For most of the myriad proteins in the postsynaptic density, however, a pharmacological approach has not been possible. The introduction of gene targeting in embryonic stem (ES) cells allowed the production of mice in which specific genes are deleted (Capecchi, 1989; Silva et al., 1992 ). This technology enabled many advances in vertebrate biology including neuroscience but suffers from two limitations. First, the gene deletion is global and in many cases lethal. Second, the deletion occurs in the germline, raising the possibility of compensation during development. The introduction of conditional knockout (KO) technology has circumvented some of these problems (Adesnik et al., 2008; Plück, 1996; Sauer and Henderson, 1988; Tsien et al., 1996 ). In this approach, the selective expression of Cre recombinase restricts gene deletion to those cells expressing Cre. This approach nonetheless requires the production of conditional KO mice, involving considerable time and expense. RNA interference (RNAi) provides a more facile way to knock down protein expression (Ehrlich and Malinow, 2004; Elias et al., 2006; Fire et al., 1998; Futai et al., 2007; Hannon, 2002 ) but has its own limitations. Off-target effects can occur (Alvarez et al., 2006; Persengiev et al., 2004 ). Although this problem can be addressed by rescuing the loss of function with an RNAi-resistant construct, the lack of effect in an animal lacking the gene is even more definitive. More important, however, RNAi rarely eliminates the protein, making it difficult to determine whether any residual function reflects residual protein or an unrelated process.

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New genome editing approaches have recently been developed that have the potential to circumvent many of the problems associated with homologous recombination in ES cells and RNAi (Wijshake et al., 2014). These include TALENs, zinc finger nucleases, and more recently clustered regularly interspaced short palindromic repeats (CRISPR). Originally discovered in bacteria as an adaptive immune defense mechanism against viral attack, CRISPR has now...