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
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.