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Full-length murine WT and 4A were BFP-tagged at the N-Terminus through cloning into the pTag-BFP-C vector (Evrogen) and PCR-cloned using Q5 High-Fidelity DNA Polymerase (NEB) and T4 DNA ligase (NEB) in-frame into the pWPI vector (Addgene) using the following primers: CDS Start + PmeI: 5′-TAAGCAGTTTAAACATGAGCGAGCTGATTAAGGAG-3′ CDS End + PmeI: 5′-TGCTTAGTTTAAACTCAGCAGTATTTCCGAGATGC- 3′ sgRNA that targeted CDK9 or no specific target (sgSCR) were cloned into the all-in-one lentiCRISPR v2 vector (Addgene), in which the selection marker puromycin was replaced by the fluorescent protein mAmetrine
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