GPCR-targeted PROTACs in Cancer Therapeutics

Molecular Pharmacology, Год журнала: 2024, Номер 107(2), С. 100013 - 100013

Опубликована: Дек. 12, 2024

G protein-coupled receptors (GPCRs) comprise a family of heptahelical membrane proteins that mediate intracellular and intercellular transmembrane signaling. Defects in GPCR signaling pathways are implicated the pathophysiology many diseases, including cardiovascular disease, endocrinopathies, immune disorders, cancer. Although GPCRs attractive drug targets, only small number Food Drug Administration-approved anticancer therapeutics target GPCRs. Targeted protein degradation (TPD) technology allows for direct modulation cellular expression level interest. TPD methods such as proteolysis-targeting chimeras (PROTACs) use ubiquitin-proteasome system to degrade interest selectively. PROTAC has not been widely applied other proteins, there is evidence PROTACs or could be GPCRome. Current show feasibility using GPCRs; however, mechanism some these uncertain. Additional studies aimed at elucidating with necessary. Discovery new allosteric molecule binders will required development intracellularly oriented PROTACs. Promising early results targeted suggest discovery platforms useful developing targeting pathological SIGNIFICANCE STATEMENT: Aberrant can contribute generally highly individual currently undrugged traditional approaches. technologies, chimeras, provide approach previously undruggable relevant molecular

Язык: Английский

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Combination of AID2 and BromoTag expands the utility of degron-based protein knockdowns

EMBO Reports, Год журнала: 2024, Номер 25(9), С. 4062 - 4077

Опубликована: Авг. 23, 2024

Язык: Английский

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Protein degradation by small tag artificial bacterial E3 ligase

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Сен. 7, 2024

ABSTRACT Targeting of proteins for degradation in a reversible manner is powerful approach to decipher gene function and mimic drug effects, with great potential target discovery validation. A generalized tag protein interest then use this recruit an endogenously or exogenously expressed E3 ligase its polyubiquitination subsequent via 26S proteasome. However, the often bulky size variability substrate-dependent efficiency mammalian ligases pose challenges practice. Here we show that small tags (10-15 amino acids) can be used efficiently endogenous when coupled rtificial b acterial E 3 (ABEL) consisting tag-interacting moiety catalytic domain bacterial IpaH9.8. We name versatile efficient platform de gradation by s mall ABEL (DESTABEL). Furthermore, containing nanobody against human α-synuclein mediates primary neurons as well adult mouse brain. Taken together, our data tag-dependent independent ABELs are yet flexible tools studies

Язык: Английский

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High-throughput optimized prime editing mediated endogenous protein tagging for pooled imaging of protein localization

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Сен. 17, 2024

Abstract The subcellular organization of proteins carries important information on cellular state and gene function, yet currently there are no technologies that enable accurate measurement protein localizations at scale. Here we develop an approach for pooled endogenous tagging using prime editing, which coupled with optical readout sequencing, provides a snapshot proteome in manner akin to perturbation-based CRISPR screens. We constructed library 17,280 pegRNAs designed exhaustively tag 60 spanning diverse localization patterns explore large space genomic pegRNA design parameters. Pooled measurements efficiency uncovered both features associated increased efficiency, including epigenetic states interactions transcription. integrate into computational model predictive value constrain the large-scale peptide knock-in. Lastly, show combining in-situ sequencing high-throughput deep learning image analysis, enables exploration many parallel following single lentiviral transduction, setting stage scalable studies dynamics across cell types environmental perturbations.

Язык: Английский

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Degron-modified Cas12a enhances single-cell CRISPR screening

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Дек. 8, 2024

Abstract Single-cell CRISPR (Perturb-seq) screens have primarily relied on Cas9 whereas Cas12a, despite its unique effectiveness for multiplex guide expression, remains underexplored. This may be due to Cas12a’s RNA array (pre-crRNA) self-processing activity and the subsequent challenges associated with pre-crRNA sequence recovery. By developing modified expression vectors a degron-based Cas12a system, we overcome constraint, allowing accurate detection of pre-crRNAs at single-cell level, thus greatly expanding possibilities future Perturb-seq efforts.

Язык: Английский

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