The effector molecule is a short RNA duplex, composed of two strands of approximately 21 nucleotides, with two overhanging bases at the 3’ end (
Elbashir et al., 2001). However, several types of precursors can trigger gene silencing, such as long double-stranded RNAs (dsRNAs) from 200 to 800 base pairs (bp) (
Guo et al., 2018), hairpin RNAs (hpRNAs) (
Hussain et al., 2019), small interfering RNAs (siRNAs) (
Guo et al., 2018), short hairpin RNAs (shRNAs) (
Wang et al., 2018), among others. Notoriously, an ingenious and inexpensive way to produce shRNAs in large quantities was developed, via
in vitro transcription of a circularized DNA template, generating multimeric shRNAs (mshRNAs) (
Seyhan et al., 2006;
Abe et al., 2012). In the last decade, such multimeric shRNAs generated by rolling circle transcription have been used by other groups and have been shown to be an interesting option for RNAi (
Wang et al., 2015;
Shopsowitz et al., 2016;
Wu et al., 2016;
Kim et al., 2018).
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