biostudies-literatureCui XBreast Cancer Research Foundation of AlabamaCongressionally Directed Medical Research ProgramsNCI NIH HHS38https://www.ebi.ac.uk/biostudies/studies/S-EPMC8819949biostudies-literatureUnknown21(1)<h4>Background</h4>Unlike autosomal tumor suppressors, X-linked tumor suppressors can be inactivated by a single hit due to X-chromosome inactivation (XCI). Here, we argue that targeted reactivation of the non-mutated allele from XCI offers a potential therapy for female breast cancers.<h4>Methods</h4>Towards this goal, we developed a dual CRISPR interference and activation (CRISPRi/a) approach for simultaneously silencing and reactivating multiple X-linked genes using two orthogonal, nuclease-deficient CRISPR/Cas9 (dCas9) proteins.<h4>Results</h4>Using Streptococcus pyogenes dCas9-KRAB for silencing XIST and Staphylococcus aureus dCas9-VPR for activating FOXP3, we achieved CRISPR activation of FOXP3 in various cell lines of human female breast cancers. In human breast cancer HCC202 cells,Molecular cancerDual CRISPR interference and activation for targeted reactivation of X-linked endogenous FOXP3 in human breast cancer cells.PMC8819949P30 CA013148W81XWH-17-1-0018W81XWH-17-1-0017Li XStringer-Reasor EWang SBae SZeng LCui XLiu RQi LSZhao DZhang CXu ZWang LfalseDual CRISPR interference and activation for targeted reactivation of X-linked endogenous FOXP3 in human breast cancer cells.<h4>Background</h4>Unlike autosomal tumor suppressors, X-linked tumor suppressors can be inactivated by a single hit due to X-chromosome inactivation (XCI). Here, we argue that targeted reactivation of the non-mutated allele from XCI offers a potential therapy for female breast cancers.<h4>Methods</h4>Towards this goal, we developed a dual CRISPR interference and activation (CRISPRi/a) approach for simultaneously silencing and reactivating multiple X-linked genes using two orthogonal, nuclease-deficient CRISPR/Cas9 (dCas9) proteins.<h4>Results</h4>Using Streptococcus pyogenes dCas9-KRAB for silencing XIST and Staphylococcus aureus dCas9-VPR for activating FOXP3, we achieved CRISPR activation of FOXP3 in various cell lines of human female breast cancers. In human breast cancer HCC202 cells,2022-01-01T00:00:00Z2022 Feb2025-04-18T16:34:09.128Z2024-11-08T11:42:42.996ZS-EPMC88199493513092510.1186/s12943-021-01472-x