Protocol detail

Activating Endogenous CDK13 using CRISPR-Cas9

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In order to activate endogenous CDK13 we used a CRISPR-Cas9 complex with three SAM components: dCas9–VP64 (Addgene #61425),MS2–p65–HSF1(Addgene, #61426), and sgRNA (Addgene, #89493) as described previously [31 (link)]. gRNAs were designed using the online optimized CRISPR design tool (http://crispr.mit.edu) and targeted the proximal promoter regions of CDK13. Oligos, synthesized by Sangon Biotech., (Shanghai, China), were annealed and sub-cloned into the lentiGuide-puro vector.

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Qi J.C., Yang Z., Lin T., Ma L., Wang Y.X., Zhang Y., Gao C.C., Liu K.L., Li W., Zhao A.N., Shi B., Zhang H., Wang D.D., Wang X.L., Wen J.K, & Qu C.B. (2021). CDK13 upregulation-induced formation of the positive feedback loop among circCDK13, miR-212-5p/miR-449a and E2F5 contributes to prostate carcinogenesis. Journal of Experimental & Clinical Cancer Research : CR, 40, 2.

Publication 2021

dCas9–VP64 MS2–p65–HSF1 sgRNA

Crispr Oligos Sam complex Vector

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Variable analysis

independent variables

CRISPR-Cas9 complex with three SAM components: dCas9–VP64 (Addgene #61425), MS2–p65–HSF1 (Addgene, #61426), and sgRNA (Addgene, #89493)

dependent variables

Activation of endogenous CDK13

control variables

Not explicitly mentioned

controls

Positive control: Not mentioned
Negative control: Not mentioned

Annotations

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