دورية أكاديمية
أعرض في EDS

Precise measurement of molecular phenotypes with barcode-based CRISPRi systems.

التفاصيل البيبلوغرافية
العنوان: Precise measurement of molecular phenotypes with barcode-based CRISPRi systems.
المؤلفون: Lobel JH; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA., Ingolia NT; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.; Lead contact.
المصدر: BioRxiv : the preprint server for biology [bioRxiv] 2024 Jun 22. Date of Electronic Publication: 2024 Jun 22.
نوع المنشور: Journal Article; Preprint
اللغة: English
بيانات الدورية: Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet ISSN: 2692-8205 (Electronic) Linking ISSN: 26928205 NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE
مستخلص: Genome-wide CRISPR-Cas9 screens have untangled regulatory networks and revealed the genetic underpinnings of diverse biological processes. Their success relies on experimental designs that interrogate specific molecular phenotypes and distinguish key regulators from background effects. Here, we realize these goals with a generalizable platform for CRISPR interference with barcoded expression reporter sequencing (CiBER-seq) that dramatically improves the sensitivity and scope of genome-wide screens. We systematically address technical factors that distort phenotypic measurements by normalizing expression reporters against closely-matched control promoters, integrated together into the genome at single copy. To test our ability to capture post-transcriptional and post-translational regulation through sequencing, we screened for genes that affected nonsense-mediated mRNA decay and Doa10-mediated cytosolic protein decay. Our optimized CiBER-seq screens accurately capture the known components of well-studied RNA and protein quality control pathways with minimal background. These results demonstrate the precision and versatility of CiBER-seq for dissecting the genetic networks controlling cellular behaviors.
Competing Interests: Declaration of interests: N.T.I holds equity and serves as a scientific advisor to Tevard Biosciences, and holds equity in Velia Therapeutics.
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معلومات مُعتمدة: F32 GM148044 United States GM NIGMS NIH HHS; R01 GM135233 United States GM NIGMS NIH HHS; S10 OD028511 United States OD NIH HHS
تواريخ الأحداث: Date Created: 20240701 Latest Revision: 20240709
رمز التحديث: 20240709
مُعرف محوري في PubMed: PMC11213135
DOI: 10.1101/2024.06.21.600132
PMID: 38948701
قاعدة البيانات: MEDLINE
الوصف
تدمد:2692-8205
DOI:10.1101/2024.06.21.600132