The clustered regularly interspaced short palindromic
repeats-CRISPR-associated protein 9 (CRISPR-Cas9) system is considered as a
technological revolution in targeted mutagenesis. However, a large amount of
time and cost is needed to screen for the CRISPR-Cas9 induced mutants from a
usual large number of initial samples. Thus, Chun Wang and Kejian Wang from
the Chinese Academy of Agricultural Sciences presented a cost-effective and
sensitive screening technique for identifying mutants based on conventional
polymerase chain reaction (PCR). They called this new technique as annealing
at critical temperature PCR (ACT-PCR).

ACT-PCR needs only one PCR step and then execution of agarose gel
electrophoresis. Because of its simplicity, ACT-PCR is suitable for rapid,
large-scale screening of CRISPR-Cas9-induced mutants.

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