Engineered genetic circuits for mammalian cells often require extensive fine-tuning to perform as intended. We present a robust, general, scalable system, called 'Boolean logic and arithmetic through DNA excision' (BLADE), to engineer genetic circuits with multiple inputs and outputs in mammalian cells with minimal optimization. The reliability of BLADE arises from its reliance on recombinases under the control of a single promoter, which integrates circuit signals on a single transcriptional layer. We used BLADE to build 113 circuits in human embryonic kidney and Jurkat T cells and devised a quantitative, vector-proximity metric to evaluate their performance. Of 113 circuits analyzed, 109 functioned (96.5%) as intended without optimization. The circuits, which are available through Addgene, include a 3-input, two-output full adder; a 6-input, one-output Boolean logic look-up table; circuits with small-molecule-inducible control; and circuits that incorporate CRISPR-Cas9 to regulate endogenous genes. BLADE enables execution of sophisticated cellular computation in mammalian cells, with applications in cell and tissue engineering.
Characterization of the C-terminal DNA-binding/DNA endonuclease region of a group II intron-encoded protein
Expanding the zinc-finger recombinase repertoire: directed evolution and mutational analysis of serine recombinase specificity determinants
Unique nucleotide sequence-guided assembly of repetitive DNA parts for synthetic biology applications
A next-generation dual-recombinase system for time- and host-specific targeting of pancreatic cancer
Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance
Detection of pathological biomarkers in human clinical samples via amplifying genetic switches and logic gates
Retrohoming of a Mobile Group II Intron in Human Cells Suggests How Eukaryotes Limit Group II Intron Proliferation
Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity
In vivo genome editing in animals using AAV-CRISPR system: applications to translational research of human disease
Linear double-stranded DNAs as innovative biological parts to implement genetic circuits in mammalian cells
Synthesizing AND gate minigene circuits based on CRISPReader for identification of bladder cancer cells
A mechanistic model of the BLADE platform predicts performance characteristics of 256 different synthetic DNA recombination circuits.
Cell Fate Conversion By mRNA
mRNA-based technology is being studied as a potential technology that could be used to reprogram cell fate. This technique provides the potential to generate safe reprogrammed cells that can be used for clinical applications. Here is the latest research on cell fate conversion by mRNA.
CRISPR for Genome Editing
Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.
CRISPR Ribonucleases Deactivation
CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.
Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.