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. Author manuscript; available in PMC: 2021 Jan 25.
Published in final edited form as: N Biotechnol. 2019 Aug 16;54:28–33. doi: 10.1016/j.nbt.2019.08.004

Figure 2:

Figure 2:

Verification of miR-22 overexpression and HIPK1 knockout

A) Real time qPCR of reverse-transcribed total cellular mRNA, using primers targeting precursor miR-22 and mature miR-22-3p confirm increased transcription levels of miR-22 in Luc-HEK-miR-22. RNU6B was used as an endogenous control. B) Clustal Omega DNA sequence alignment of CRISPR HIPK1 guide RNA target and surrounding regions, showing mutations in the CRISPR/CAS9 treated cell line Luc-HEK-HIPK1-KO (3-1-2-8) compared to the parental Luc-HEK cell line. Highlighted region is the gRNA target sequence. C) Clustal Omega DNA sequence alignment from 10 colonies of TOPO cloned Luc-HEK-HIPK1 cells demonstrate that the mutation is biallelic.