IP Library › Granted Patent US 12,428,633
Granted Patent B2
US 12,428,633 · App. 15/838,598 · Granted Sep 30, 2025

Inducible dimerization of recombinases

Inventors: Wilson W. Wong (Brookline, MA); Benjamin Harris Weinberg (Boston, MA)
Assignee: TRUSTEES OF BOSTON UNIVERSITY
C12N9/96C12N9/00
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Quick Facts
Patent No.
US 12,428,633
App. No.
15/838,598
Granted
Sep 30, 2025
Kind
B2
Abstract

The technology described herein relates to controlled chemically- or light-induced rejoinder of split-recombinases. In some embodiments, compositions, methods, kits and systems are provided that relate to a split-recombinase system, whereby protein complementation or rejoinder of split-recombinases is mediated by chemical-induced dimerization domains (CIDDs) or light-induced dimerization domains (LIDD) and rejoinder of the split-recombinases occur in the presence of one or more chemical inducers or light inducers, respectively. The split-recombinases systems as disclosed herein can be used in gene therapy, integrated logic and memory in living cells such as mammalian cells. The nucleic acid cassettes, switches, and systems described herein allow for controlled gene expression or gene regulation. The controlled chemically- or light-induced rejoinder of split-recombinases can be used in, for example, adopted T-cell therapy.

Claims (47)

1. A split-recombinase polypeptide, comprising:

a. at least two complementary protein pairs (CPP), wherein each CPP comprises a binding motif (CPP A ) and a complementary binding motif (CPP B ), wherein each CPP A and CPP B are a pair of chemically-induced dimerization domains (CIDDs), or wherein each CPP A and CPP B are a pair of light-induced dimerization domains (LIDDs), and wherein the at least two CPPs comprise:

i. at least a first CPP (CPP1) comprising a CPP1 binding motif (CPP1 A ) and a complementary CPP1 binding motif (CPP1 B ), wherein CPP1 A and CPP1 B are a pair of CIDDs, and CPP1 A binds to a first target agent, and CPP1 B also binds to the same first target agent, or wherein CPP1 A and CPP1 B are a pair of LIDDs that bind to each other upon exposure to a first light signal of appropriate wavelength, and

ii. a second CPP (CPP2) comprising a CPP2 binding motif (CPP2 A ) and a complementary CPP2 binding motif (CPP2 B ), wherein CPP2 A and CPP2 B are a pair of CIDDs, and CPP2 A binds to a second target agent, and CPP2 B also binds to the same second target agent, or wherein CPP2 A and CPP2 B are a pair of LIDDs that bind to each other upon exposure to a second light signal of appropriate wavelength, and

b. a recombinase protein, wherein the recombinase protein is split into three fragments; a first recombinase polypeptide fragment (R 1 ), a second recombinase polypeptide fragment (R 2 ), and a third recombinase polypeptide fragment (R 3 ), wherein R 1 , R 2 and R 3 recombinase polypeptide fragments are each not active by themselves, and wherein R 1 , R 2 and R 3 recombinase fragments can recombine in real-time to form the recombinase protein in an active configuration;

wherein:

R 1 is conjugated to the CPP1 A ,

R 2 is conjugated to the CPP1 B and the CPP2 A , and

R 3 is conjugated to the CPP2 B ,

wherein in the presence of the first target agent, CPP1 A and CPP1 B both bind to the first target agent resulting in protein complementation of R 1 and R 2 , or upon exposure to the first light signal CPP1 A and CPP1 B bind to each other resulting in protein complementation of R 1 and R 2 , and

wherein in the presence of the second target agent, CPP2 A and CPP2 B both bind to the second target agent resulting in protein complementation of R 2 and R 3 , or upon exposure to the second light signal CPP2 and CPP2 B bind to each other resulting in protein complementation of R 2 and R 3 ,

wherein the recombinase protein is formed into the active configuration only in the presence of both (1) the first target agent or the first light signal; and (2) the second target agent or the second light signal,

wherein the recombinase protein is selected from the group consisting of: Flp, PhiC31 (@C31), SEQ ID NO: 16 or a polypeptide that has at least 85% identity to the amino acid sequence of SEQ ID NO: 16, B3, Bxb1, Dre, Vika, and FlpO, and

wherein the binding motif (CPP A ) and complementary binding motif (CPP B ) of each CPP is selected from any one of:

a. the CPP A comprises a GID1 domain or a fragment thereof, and the CPP B comprises a GAI domain, wherein the GID1 domain and GAI domain bind to the target agent Gibberellin Ester (GIB);

b. the CPP A comprises a FKBP domain or a fragment thereof, and the CPP B comprises an FRB domain, wherein the FKBP domain and FRB domain bind to the target agent Rapalog (RAP);

c. the CPP A comprises a PYL domain or a fragment thereof, and the CPP B comprises an ABI domain, wherein the PLY domain and ABI domain bind to the target agent Ascorbic-acid (ABA); or

d. the CPP A comprises a LIDD, and the CPP B comprises a complementary LIDD, wherein the LIDD dimerizes with the complementary LIDD upon exposure to a light signal of an appropriate wavelength.

2. The split-recombinase polypeptide of claim 1 , wherein the first (R 1 ), second (R 2 ) and third (R 3 ) recombinase polypeptide fragments are Flp recombinase polypeptide fragments, wherein R 1 , R 2 and R 3 are selected from any of:

a. R 1 comprises amino acids 1-27 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 28-168 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 169-423 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto;

b. R 1 comprises amino acids 1-27 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 28-374 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 375-423 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto;

c. R 1 comprises amino acids 1-27 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 28-396 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 397-423 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto;

d. R 1 comprises amino acids 1-168 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 169-374 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 375-423 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto;

e. R 1 comprises amino acids 1-168 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 169-396 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 397-423 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and

f. R 1 comprises amino acids 1-374 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 375-396 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 397-423 of SEQ ID NO: 1 or a polypeptide that has 85% sequence identity thereto.

3. The split-recombinase polypeptide of claim 1 , wherein the first (R 1 ), second (R 2 ) and third (R 3 ) recombinase polypeptide fragments are PhiC31 recombinase polypeptide fragments, wherein R 1 , R 2 and R 3 are selected from any of:

a. R 1 comprises amino acids 1-233 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 234-369 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 397-605 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto;

b. R 1 comprises amino acids 1-233 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 234-428 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 429-605 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto;

c. R 1 comprises amino acids 1-233 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 234-571 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 572-605 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto;

d. R 1 comprises amino acids 1-396 of SEQ ID NO: 12 or a polypeptide that is has 85% sequence identity thereto; and R 2 comprises amino acids 397-428 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 429-605 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto;

e. R 1 comprises amino acids 1-396 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 397-571 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 572-605 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and

f. R 1 comprises amino acids 1-428 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 429-571 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 572-605 of SEQ ID NO: 12 or a polypeptide that has 85% sequence identity thereto.

4. The split-recombinase polypeptide of claim 1 , wherein the first (R 1 ), second (R 2 ) and third (R 3 ) recombinase polypeptide fragments are VCre recombinase polypeptide fragments, wherein R 1 , R 2 and R 3 are selected from any of:

a. R 1 comprises amino acids 1-82 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 83-172 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 173-380 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto;

b. R 1 comprises amino acids 1-82 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 83-269 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 270-380 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto;

c. R 1 comprises amino acids 1-82 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 83-277 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 278-380 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto;

d. R 1 comprises amino acids 1-172 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 173-269 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 270-380 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto;

e. R 1 comprises amino acids 1-172 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 173-277 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 278-380 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and

f. R 1 comprises amino acids 1-269 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 2 comprises amino acids 270-277 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto; and R 3 comprises amino acids 278-380 of SEQ ID NO: 16 or a polypeptide that has 85% sequence identity thereto.

5. The split-recombinase polypeptide of claim 1 , wherein the LIDD is nMag or CIBN, wherein nMag dimerizes with the complementary LIDD pMag upon exposure to a blue light signal, and wherein CIBN dimerizes with the complementary CRY2 upon exposure to a blue light signal.

6. The split-recombinase polypeptide of claim 5 , wherein the blue light signal is a pulse light signal.

7. The split-recombinase of claim 1 , wherein the active recombinase protein can recognize the recombinant recognition sequence (RRS) of the recombinase protein.

8. The split-recombinase polypeptide of claim 1 , wherein the GID1 domain comprises an amino acid sequence of SEQ ID NO: 31 or a polypeptide with 85% sequence identity to SEQ ID NO: 31, and the GAI domain comprises an amino acid sequence of SEQ ID NO: 30 or a polypeptide with 85% sequence identity to SEQ ID NO: 30.

9. The split-recombinase polypeptide of claim 1 , wherein the FKBP domain comprises an amino acid sequence of SEQ ID NO: 22 or a polypeptide with 85% sequence identity to SEQ ID NO: 22, and the FBP domain comprises an amino acid sequence of SEQ ID NO: 23 or a polypeptide with 85% sequence identity to SEQ ID NO: 23.

10. The split-recombinase polypeptide of claim 1 , wherein the PYL domain comprises an amino acid sequence of SEQ ID NO: 26 or a polypeptide with 85% sequence identity to SEQ ID NO: 26, and the ABI domain comprises an amino acid sequence of SEQ ID NO: 27 or a polypeptide with 85% sequence identity to SEQ ID NO: 27.

11. The split-recombinase polypeptide of claim 5 , wherein the nMag LIDD comprises an amino acid sequence of SEQ ID NO: 51 or a polypeptide with 85% sequence identity to SEQ ID NO: 51, and the pMag complementary LIDD comprises an amino acid sequence of SEQ ID NO: 52 or a polypeptide with 85% sequence identity to SEQ ID NO: 52.

12. The split-recombinase polypeptide of claim 5 , wherein the CIBN LIDD comprises an amino acid sequence of SEQ ID NO: 55 or a polypeptide with 85% sequence identity to SEQ ID NO: 55, and the CRY2 complementary LIDD comprises an amino acid sequence of SEQ ID NO: 56 or a polypeptide with 85% sequence identity to SEQ ID NO: 56.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 22, 2018
From: WONG, WILSON W.; WEINBERG, BENJAMIN HARRIS
To: TRUSTEES OF BOSTON UNIVERSITY
Reel/Frame 045001/0780 →
Continuity (2)
Provisional Application 62432916 · Dec 12, 2016
Related Publication 20180163195A1 · Jun 14, 2018
References Cited (78)
US 20090031441A1 · Matsuoka et al. · 2009 [cited by applicant]
US 20120003630A1 · Collins · 2012 [cited by applicant]
US 20140315310A1 · Lu · 2014 [cited by applicant]
US 20160264665A1 · Lim et al. · 2016 [cited by applicant]
US 20170306336A1 · Lu · 2017 [cited by examiner]
WO 2015188191A1 · 2015 [cited by applicant]
WO WO2015188094A1 · 2015 [cited by examiner]
Guo et al., Nature vol. 389, pp. 40-46, Sep. 1997. [cited by examiner]
Roybal et al., “Precision Tumor Recognition by T Cells With Combinatorial Antigen-Sensing Circuits”, Cell 164(4) 770-779 (2016). [cited by applicant]
Sajgo et al., “Dre-Cre sequential recombination provides new tools for retinal ganglion cell labeling and manipulation in mice”, PLoS One 9(3) e91435 (2014). [cited by applicant]
Sarkar et al., “HIV-1 proviral DNA excision using an evolved recombinase”, Science 316(5833) 1912-1915 (2007). [cited by applicant]
Sauer et al., “DNA recombination with a heterospecific Cre homolog identified from comparison of the pac-c1 regions of P1-related phages”, Nuclec Acids 32(20) 6086-6095 (2004). [cited by applicant]
Schonhuber et al., “A next-generation dual-recombinase system for time- and host-specific targeting of pancreatic cancer”, Nat Med 20(11) 1340-1347 (2014). [cited by applicant]
Shaikh et al., “Chimeras of the Flp and Cre recombinases: tests of the mode of cleavage by Flp and Cre”, J Mol Biol 302(1) 27-48 (2000). [cited by applicant]
Shannon et al., “The Synthesis of Two-Terminal Switching Circuits”, Bell Labs Technical Journal 28(1) 59-98 (1949). [cited by applicant]
Sirk et al., “Expanding the zinc-finger recombinase repertoire: directed evolution and mutational analysis of serine recombinase specificity determinants”, Nucleic Acids Res 42(7) 4755-4766 (2014). [cited by applicant]
Situti et al., “Synthetic circuits integrating logic and memory in living cells”, Nat Biotechnol 31(5) 448-452 (2013). [cited by applicant]
Slomovic et al., “DNA sense-and-respond protein modules for mammalian cells”, Nat Methods 12(11) 1085-1090 (2015). [cited by applicant]
Stanton et al., “Genomic mining of prokaryotic repressors for orthogonal logic gates”, Nat Chem Biol 10(2) 99-105 (2014). [cited by applicant]
Stricker et al., “A fast, robust and tunable synthetic gene oscillator”, Nature 456(7221) 516-519 (2008). [cited by applicant]
Suzuki et al., “VCre/VloxP and SCre/SloxP: new site-specific recombination systems for genome engineering”, Nucleic Acids Res 39(8) e49 (2011). [cited by applicant]
Tamsir et al., “Robust multicellular computing using genetically encoded NOR gates and chemical ‘wires’”, Nature 469(7329) 212-215 (2011). [cited by applicant]
Torella et al., “Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly”, Nucleic Acids Res 42(1) 681-689 (2014). [cited by applicant]
Torella et al., “Unique nucleotide sequence-guided assembly of repetitive DNA parts for synthetic biology applications”, Nat Protoc 9(9) 2075-2089 (2014). [cited by applicant]
Torikai et al., “Toward eliminating HLA class I expression to generate universal cells from allogeneic donors”, Blood 122: 1341-1349 (2013). [cited by applicant]
Weber et al., “A synthetic time-delay circuit in mammalian cells and mice”, Proc Natl Acad Sci 104(8) 2643-2648 (2007). [cited by applicant]
Wei et al., “Bacterial virulence proteins as tools to rewire kinase pathways in yeast and immune cells”, Nature 488 (7411) 384-388 (2012). [cited by applicant]
Xie et al., “Multi-input RNAi-based logic circuit for identification of specific cancer cells”, Science 333(6047) 1307-1311 (2011). [cited by applicant]
Zhao et al., “Heterelogous expression of mutated HLA-G decreases immunogenicity of human embryonic stem cells and their epidermal derivatives”, Stem Cell Res 13(2) 342-354 (2014). [cited by applicant]
Akopian et al., “Chimeric recombinases with designed DNA sequence recognition”, Proc Natl Acad Sci USA 100(15) 8688-8691 (2003). [cited by applicant]
Appleton et al.,+55:78 “Interactive assembly algorithms for molecular cloning”, Nat Methods 11(6) 657-662 (2014). [cited by applicant]
Blomfield et al., “The regulation of pap and type 1 fimbriation in Escherichia coli”, Adv Microb Physiol 45: 1-49 (2001). [cited by applicant]
Bogorad et al., “Synthetic non-oxidative glycolysis enables complete carbon conservation”, Nature 502(7473) 693-697 (2013). [cited by applicant]
Bonnet et al., “Amplifying genetic logic gates”, Science 340(6132) 599-603 (2013). [cited by applicant]
Bonnet et al., “Rewritable digital data storage in live cells via engineered control of recombination directionality”, Proc Natl Acad Sci USA 109(23) 8884-8889 (2012). [cited by applicant]
Branda et al., “Talking about a revolution: The impact of site-specific recombinases on genetic analyses in mice”, Dev Cell 6(1) 7-28 (2004). [cited by applicant]
Brophy et al., “Principles of genetic circuit design”, Nat Methods 11(5) 508-520 (2014). [cited by applicant]
Canton et al., “Refinement and standardization of synthetic biological parts and devices”, Nat Biotechnol 26(7) 787-793 (2008). [cited by applicant]
Chakravarti et al., “Synthetic biology in cell-based cancer immunotherapy”, Trends Biotechnol 33(8) 449-461 (2015). [cited by applicant]
Chavez et al., “Highly efficient Cas9-mediated transcriptional programming”, Nat Methods 12(4) 326-328 (2015). [cited by applicant]
Chen et al., “Characterization of 582 natural and synthetic terminators and quantification of their design constraints”, Nat Methods 10(7) 659-664 (2013). [cited by applicant]
Chen et al., “Synthetic Biology. Emergent genetic oscillations in a synthetic microbial consortium”, Science 349 (6251) 986-989 (2015). [cited by applicant]
Courbert et al., “Detection of pathological biomarkers in human clinical samples via amplifying genetic switches and logic gates”, Sci Transl Med 7(289) 289ra83 (2015). [cited by applicant]
Elowitz et al., “A synthetic oscillatory network of transcriptional regulators”, Nature 403(6767) 335-338 (2000). [cited by applicant]
Farruggio et al., “Serine integrase chimeras with activity in [cited by applicant]
Fenno et al., “Targeting cells with single vectors using multiple-feature Boolean logic”, Nat Methods 11(7) 763-772 (2014). [cited by applicant]
Friedland et al., “Synthetic gene networks that count”, Science 324(5931) 1199-1202 (2009). [cited by applicant]
Gaber et al., “Designable DNA-binding domains enable construction of logic circuits in mammalian cells”, Nat Chem Biol 10(3) 203-208 (2014). [cited by applicant]
Gardner et al., “Construction of a genetic toggle switch in Escherichia coli”, Nature 403(6767) 339-342 (2000). [cited by applicant]
Gordley et al., “Synthesis of programmable integrases”, Proc Natl Acad Sci USA 106(13) 5053-5058 (2009). [cited by applicant]
Green et al., “Toehold switches: de-novo-designed regulators of gene expression”, Cell 159(4) 925-939 (2014). [cited by applicant]
Grupp et al., “Chimeric antigen receptor-modified T cells for acute lymphoid leukemia”, N Engl J Med 368(16) 1509-1518 (2013). [cited by applicant]
Guilinger et al., “Fusion of catalytically inactive Cas9 to Fokl nuclease improves the specificity of genome modification”, Nature Biotechnology 32; 577-582 (2014). [cited by applicant]
Guinn et al., “Biological 2-input decoder circuit in human cells”, ACS Synth Biol 3(8) 627-633 (2014). [cited by applicant]
Ham et al., “Design and construction of a double inversion recombination switch for heritable sequential genetic memory”, PLoS One 3(7) e2815 (2008). [cited by applicant]
Hauber et al., “Highly significant antiviral activity of HIV-1 LTR-specific tre-recombinase in humanized mice”, PLoS Pathology 9(9) e1003587 (2013). [cited by applicant]
Hsiao et al., “A population-based temporal logic gate for timing and recording chemical events”, Mol Syste Biol 12(5) 869 (2016). [cited by applicant]
Huynh et al., “Automatic design of synthetic gene circuits through mixed integer non-linear programming”, PLoS One 7(4) e35529 (2012). [cited by applicant]
Jayanthi et al., “Retroactivity controls the temporal dynamics of gene transcription”, ACS Synth Biol 2(8) 431-441 (2013). [cited by applicant]
Johnson, “Chapter 13: Bacterial Site-Specific SNA Inversion Systems” Mobile DNA II, ASM Press, Washington, D.C., 2002. [cited by applicant]
Jullien et al., “Regulation of Cre recombinase by ligand-induced complementation of inactive fragments”, Nucleic Acids Res 31(21) e131 (2003). [cited by applicant]
Karimova et al., “Vika/vox, a novel efficient and specific Cre/loxP-like site-specific recombination system”, Nucleic Acids Res 41(2) e37 (2013). [cited by applicant]
Karpinski et al., “Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity”, Nat Biotechnol 34(4) 401-409 (2016). [cited by applicant]
Khalil et al., “A synthetic biology framework for programming eukaryotic transcription functions”, Cell 150(3) 647-658 (2012). [cited by applicant]
Khalil et al., “Synthetic biology: applications come of age”, Nat Rev Genet 11(5) 367-379 (2010). [cited by applicant]
Lee et al., “Role of nucleotide sequences of loxP spacer region in Cre-mediated recombination”, Gene 216(1) 55-65 (1998). [cited by applicant]
Leisner et al., “Rationally designed logic integration of regulatory signals in mammalian cells”, Nat Nanotechnol 5(9) 666-667 (2010). [cited by applicant]
Madisen et al., “Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance”, Neuron 85(5) 942-958 (2015). [cited by applicant]
Mercer et al., “Chimeric TALE recombinases with programmable DNA sequence specificity”, Nucleic Acids Res 40 (21) 11163-11172 (2012). [cited by applicant]
Moon et al., “Genetic programs constructed from layered logic gates in single cells”, Nature 491(7423) 249-253 (2012). [cited by applicant]
Morgan et al., “Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2”, Mol Ther 18(4) 843-851 (2010). [cited by applicant]
Mutalik et al., “Quantitative estimation of activity and quality for collections of functional genetic elements”, Nat Methods 10(4) 347-353 (2013). [cited by applicant]
Nagy, “Cre recombinase: the universal reagent for genome tailoring”, Genesis 26(2) 99-109 (2000). [cited by applicant]
Neilsen et al., “Genetic circuit design automation”, Science 352(6281) aac7341 (2016). [cited by applicant]
Regot et al., “Distributed biological computation with multicellular engineered networks”, Nature 469(7329) 207-211 (2011). [cited by applicant]
Ro et al., “Production of the antimalarial drug precursor artemisinic acid in engineered yeast”, Nature 440(7086) 940-943 (2006). [cited by applicant]
Rodrigo et al., “AutoBioCAD: full biodesign automation of genetic circuits”, ACS Synth Biol 2(5) 230-236 (2013). [cited by applicant]
Roquet et al., “Synthetic recombinase-based state machines in living cells”, Science 353(6297) aad8559 (2016). [cited by applicant]