Scalable recombinase-based gene expression cascades

Autor: Benjamin H. Weinberg, Timothy K. Lu, Tackhoon Kim, Wilson W. Wong
Rok vydání: 2021
Předmět:
0301 basic medicine
Transcription
Genetic
Carcinogenesis
Computer science
Science
Cellular differentiation
Gene regulatory network
Transposases
General Physics and Astronomy
Locus (genetics)
Computational biology
medicine.disease_cause
ENCODE
General Biochemistry
Genetics and Molecular Biology
03 medical and health sciences
Synthetic biology
0302 clinical medicine
CRISPR-Associated Protein 9
Gene expression
medicine
Recombinase
Humans
Gene Regulatory Networks
Cellular programming
Gene
Gene Editing
Mutation
Multidisciplinary
Genome
Human
High-Throughput Nucleotide Sequencing
Cell Differentiation
General Chemistry
HEK293 Cells
ComputingMethodologies_PATTERNRECOGNITION
030104 developmental biology
Genetic Loci
Scalability
ComputingMethodologies_GENERAL
CRISPR-Cas Systems
Genetic Engineering
030217 neurology & neurosurgery
Plasmids
RNA
Guide
Kinetoplastida
Zdroj: Nature Communications, Vol 12, Iss 1, Pp 1-9 (2021)
ISSN: 2041-1723
DOI: 10.1038/s41467-021-22978-4
Popis: Temporal modulation of the expression of multiple genes underlies complex complex biological phenomena. However, there are few scalable and generalizable gene circuit architectures for the programming of sequential genetic perturbations. Here, we describe a modular recombinase-based gene circuit architecture, comprising tandem gene perturbation cassettes (GPCs), that enables the sequential expression of multiple genes in a defined temporal order by alternating treatment with just two orthogonal ligands. We use tandem GPCs to sequentially express single-guide RNAs to encode transcriptional cascades that trigger the sequential accumulation of mutations. We build an all-in-one gene circuit that sequentially edits genomic loci, synchronizes cells at a specific stage within a gene expression cascade, and deletes itself for safety. Tandem GPCs offer a multi-tiered cellular programming tool for modeling multi-stage genetic changes, such as tumorigenesis and cellular differentiation. There are few robust circuit architectures for sequential gene perturbations. Here, the authors use a modular recombinase-based design that sequentially edits loci, synchronizes cells, and deletes itself.
Databáze: OpenAIRE
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