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miR-KO System

Disrupt miR genes with
TALE Nucleases and HR vectors

Knockout microRNAs on the genomic level

  • Pre-designed TALE Nucleases pairs
  • Matched HR donor vectors
  • Easy selection of miR-KO clonal lines
  • Verify knockout by qPCR
  • Discover novel miR phenotypes

Completely ablate microRNA genes on the genomic level

miR-KOs are transcription activator-like effector (TALE) nucleases that precisely edit specific miRNAs in mammalian cells. SBI designed miR-TALE-nucleases to cleave within the miRNA seed region. In the absence of HR donor vectors, the cellular machinery repairs such breaks via non-homologous end joining (NHEJ). This is an error-prone system that typically generates small deletions or insertions (indels) at or near the site of cleavage. Since the seed region (defined as bases 2-8 of the microRNA) directs miRNA binding to its target DNA, indels within the seed region completely abolish miRNA function.

Design of miR-KO TALENs

miR-KO TALEN design

The miR-KOs are designed to disrupt the miRNA seed region. Pairing miR-KOs with an HR donor replaces the entire miRNA hairpin structure with an insulated selectable marker cassette.