Granted Patent
B2
US 12,624,362 · App. 17/280,294 · Granted May 12, 2026
Mutant reverse tetracycline transactivators for expression of genes
Inventors:
David A. Sinclair (Cambridge, MA); Yuancheng Lu (Cambridge, MA); Noah Justin Davidsohn (La Jolla, CA)
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Abstract
Provided herein are mutant reverse tetracycline transactivator (rtTA) proteins and engineered nucleic acids that encode a mutant rtTA that are useful in regulating gene expression, inducing cellular reprogramming, tissue repair, tissue regeneration, organ regeneration, reversing aging, treating a disease, or any combination thereof. Also provided herein are recombinant viruses comprising the engineered nucleic acids and methods of regulating cellular reprogramming, tissue repair, tissue regeneration, or any combination thereof by administering an engineered nucleic acid or recombinant virus comprising the same in a cell, tissue or subject comprising administering a mutant rtTA and an inducible nucleic acid encoding a transgene.
Claims (37)
1 . An engineered nucleic acid comprising a promoter operably linked to a first nucleic acid comprising a sequence that is at least 80% identical to the sequence of SEQ ID NO: 12, wherein the first nucleic acid sequence encodes a mutant reverse tetracycline transactivator (rtTA) comprising an amino acid sequence at least 70% identical to the sequence of SEQ ID NO: 11, which comprises the following mutations relative to rtTA3 (SEQ ID NO: 11):
(a) a G72V mutation;
(b) a G12S or G12T mutation;
(c) an F67S or F67T mutation; and
(d) an R171K or R171H mutation.
2 . The engineered nucleic acid of claim 1 , wherein the promoter is a constitutive promoter selected from the group consisting of CP1, CMV, EF1a, SV40, PGK1, Ubc, human beta actin, CAG, Ac5, polyhedrin, TEF1, GDS, CaMV 35S, Ubi, H1, and U6 promoters.
3 . The engineered nucleic acid of claim 1 , wherein the promoter is a tissue-specific promoter.
4 . The engineered nucleic acid of claim 1 further comprising a second nucleic acid sequence that encodes a tetracycline repressor.
5 . The engineered nucleic acid of claim 4 , wherein the promoter that is operably linked to the first nucleic acid is also operably linked to the second nucleic acid.
6 . The engineered nucleic acid of claim 4 further comprising a separator sequence encoding an internal ribosome entry site (IRES) or a 2A peptide between the first and the second nucleic acid.
7 . The engineered nucleic acid of claim 4 , wherein the tetracycline repressor is tetRKRAB.
8 . The engineered nucleic acid of claim 1 , wherein the engineered nucleic acid is a viral vector, wherein the viral vector is an adeno-associated virus (AAV), lentiviral, retroviral, adenoviral, or herpes viral vector.
9 . The engineered nucleic acid of claim 8 , wherein the viral vector comprises inverted terminal repeats (ITRs) flanking one or more nucleic acids.
10 . The engineered nucleic acid of claim 1 further comprising a WPRE3 sequence.
11 . The engineered nucleic acid of claim 1 , wherein the engineered nucleic acid comprises a sequence that is at least 70% identical to the sequence of SEQ ID NO: 17 or SEQ ID NO: 30.
12 . The engineered nucleic acid of claim 1 , wherein the engineered nucleic acid further comprises a first transgene sequence operably linked to an inducible promoter that comprises a tetracycline-responsive element (TRE).
13 . The engineered nucleic acid of claim 12 , wherein the TRE promoter is a TRE3G promoter.
14 . The engineered nucleic acid of claim 12 , wherein the TRE comprises at least one Tet-O sequence set forth as SEQ ID NO: 19.
15 . A recombinant virus comprising the engineered nucleic acid of claim 1 .
16 . A pharmaceutical composition comprising the recombinant virus of claim 15 .
17 . A pharmaceutical composition comprising the engineered nucleic acid of claim 1 and a pharmaceutically acceptable excipient.
18 . An in vitro or ex vivo cell comprising the engineered nucleic acid of claim 1 .
19 . The engineered nucleic acid of claim 1 , wherein the mutant rtTA comprises the following mutations:
(a) G72V;
(b) G12S;
(c) F67S; and
(d) R171K.
20 . The engineered nucleic acid of claim 1 , wherein the mutant rtTA comprises a sequence that is at least 70% identical to the sequence of SEQ ID NO: 13.
21 . The engineered nucleic acid of claim 1 , wherein the mutant rtTA comprises SEQ ID NO: 13.
22 . The engineered nucleic acid of claim 1 , wherein the first nucleic acid comprises the sequence that is set forth in SEQ ID NO: 12.
23 . The engineered nucleic acid of claim 1 , wherein the engineered nucleic acid comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 17 or SEQ ID NO: 30.
24 . The engineered nucleic acid of claim 1 , wherein the engineered nucleic acid comprises the sequence that is set forth in SEQ ID NO: 17 or SEQ ID NO: 30.
25 . The engineered nucleic acid of claim 1 , wherein the mutant rtTA comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 13.
26 . The engineered nucleic acid of claim 1 , wherein the first nucleic acid comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 12.
27 . A method of promoting gene expression comprising administering to a cell, tissue, or subject in need thereof:
(a) a first engineered nucleic acid of claim 12 ; and
(b) a tetracycline, thereby promoting expression of the first transgene.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Sep 15, 2022
From: SINCLAIR, DAVID A.; LU, YUANCHENG; DAVIDSOHN, NOAH
To: PRESIDENT AND FELLOWS OF HARVARD COLLEGE
Reel/Frame 061099/0444 →
Continuity (2)
Provisional Application
62738894
· Sep 28, 2018
References Cited (325)
US 5399363A
· Liversidge et al.
· 1995
[cited by applicant]
US 5543158A
· Gref et al.
· 1996
[cited by applicant]
US 5552157A
· Yagi et al.
· 1996
[cited by applicant]
US 5565213A
· Nakamori et al.
· 1996
[cited by applicant]
US 5567434A
· Szoka, Jr.
· 1996
[cited by applicant]
US 5589362A
· Bujard et al.
· 1996
[cited by applicant]
US 5641515A
· Ramtoola
· 1997
[cited by applicant]
US 5650298A
· Bujard et al.
· 1997
[cited by applicant]
US 5738868A
· Shinkarenko
· 1998
[cited by applicant]
US 5741516A
· Webb et al.
· 1998
[cited by applicant]
US 5795587A
· Gao et al.
· 1998
[cited by applicant]
US 5928941A
· Lee et al.
· 1999
[cited by applicant]
US 6750015B2
· Horwitz et al.
· 2004
[cited by applicant]
US 7541446B2
· Hillen et al.
· 2009
[cited by applicant]
US 8080647B2
· Gordon-Kamm
· 2011
[cited by examiner]
US 8158415B2
· Jo et al.
· 2012
[cited by applicant]
US 8326547B2
· Liu et al.
· 2012
[cited by applicant]
US 8383364B2
· Berkhout et al.
· 2013
[cited by applicant]
US 8609373B2
· Liu et al.
· 2013
[cited by applicant]
US 8716465B2
· Rossi et al.
· 2014
[cited by applicant]
US 8748179B2
· Egusa et al.
· 2014
[cited by applicant]
US 8883506B2
· Rossi et al.
· 2014
[cited by applicant]
US 8932856B2
· Jaenisch et al.
· 2015
[cited by applicant]
US 8940536B2
· Jaenisch et al.
· 2015
[cited by applicant]
US 8957037B2
· Collard et al.
· 2015
[cited by applicant]
US 9127283B2
· Bisgrove et al.
· 2015
[cited by applicant]
US 9175268B2
· Mack
· 2015
[cited by applicant]
US 9175311B2
· Townes et al.
· 2015
[cited by applicant]
US 9228204B2
· Pulst et al.
· 2016
[cited by applicant]
US 9497943B2
· Jaenisch et al.
· 2016
[cited by applicant]
US 9580689B2
· Kikyo et al.
· 2017
[cited by applicant]
US 9644164B2
· Gieselman et al.
· 2017
[cited by applicant]
US 9695445B2
· Fusaki et al.
· 2017
[cited by applicant]
US 9850499B2
· Baylink et al.
· 2017
[cited by applicant]
US 9862930B2
· Dowdy et al.
· 2018
[cited by applicant]
US 9920333B2
· Pulst et al.
· 2018
[cited by applicant]
US RE49583E
· Berkhout et al.
· 2023
[cited by applicant]
US 20020165180A1
· Weaver
· 2002
[cited by applicant]
US 20030065157A1
· Lasek
· 2003
[cited by applicant]
US 20030138772A1
· Gao et al.
· 2003
[cited by applicant]
US 20030138799A1
· Ruppert et al.
· 2003
[cited by applicant]
US 20030165921A1
· Tang et al.
· 2003
[cited by applicant]
US 20030186281A1
· Hillen
· 2003
[cited by applicant]
US 20040038249A1
· Darteil et al.
· 2004
[cited by applicant]
US 20040219579A1
· Aziz et al.
· 2004
[cited by applicant]
US 20050064454A1
· Young et al.
· 2005
[cited by applicant]
US 20050208496A1
· Ohtani et al.
· 2005
[cited by applicant]
US 20060263774A1
· Clark et al.
· 2006
[cited by applicant]
US 20070042392A1
· Tang et al.
· 2007
[cited by applicant]
US 20070048301A1
· Bodary-Winter et al.
· 2007
[cited by applicant]
US 20070060743A1
· Tang et al.
· 2007
[cited by applicant]
US 20070072175A1
· Cooper et al.
· 2007
[cited by applicant]
US 20070161031A1
· Trinklein et al.
· 2007
[cited by applicant]
US 20080050379A1
· Young et al.
· 2008
[cited by applicant]
US 20080050393A1
· Tang et al.
· 2008
[cited by applicant]
US 20100048678A1
· Smit et al.
· 2010
[cited by applicant]
US 20100074864A1
· Achiron et al.
· 2010
[cited by applicant]
US 20100150889A1
· Townes et al.
· 2010
[cited by applicant]
US 20100190250A1
· Hu et al.
· 2010
[cited by applicant]
US 20100273220A1
· Yanik et al.
· 2010
[cited by applicant]
US 20110002940A1
· Piek et al.
· 2011
[cited by applicant]
US 20110061118A1
· Ralf et al.
· 2011
[cited by applicant]
US 20110190729A1
· Kirkland et al.
· 2011
[cited by applicant]
US 20110190730A1
· Kirkland et al.
· 2011
[cited by applicant]
US 20110258713A1
· Zhu et al.
· 2011
[cited by applicant]
US 20120095188A1
· Jo et al.
· 2012
[cited by applicant]
US 20120128655A1
· Kim et al.
· 2012
[cited by applicant]
US 20120225076A1
· Peeper et al.
· 2012
[cited by applicant]
US 20130059752A1
· Bodary-Winter et al.
· 2013
[cited by applicant]
US 20130065791A1
· Rosenthal et al.
· 2013
[cited by applicant]
US 20130130387A1
· Itskovitz-Eldor et al.
· 2013
[cited by applicant]
US 20140128277A1
· Moller et al.
· 2014
[cited by applicant]
US 20140323411A1
· Kostarelos
· 2014
[cited by applicant]
US 20150087594A1
· Edenhoffer et al.
· 2015
[cited by applicant]
US 20150133531A1
· Wiegand
· 2015
[cited by applicant]
US 20150267174A1
· Hayashi et al.
· 2015
[cited by applicant]
US 20160032393A1
· Achiron et al.
· 2016
[cited by applicant]
US 20160076000A1
· Townes et al.
· 2016
[cited by applicant]
US 20160102127A1
· Thepen et al.
· 2016
[cited by applicant]
US 20180305689A1
· Sætrom et al.
· 2018
[cited by applicant]
US 20190292250A1
· Hinderer
· 2019
[cited by examiner]
US 20210324414A1
· Weiss et al.
· 2021
[cited by applicant]
US 20250325628A1
· Sinclair et al.
· 2025
[cited by applicant]
AU 746850B2
· 2002
[cited by applicant]
CN 101302553A
· 2008
[cited by applicant]
CN 104837862A
· 2015
[cited by applicant]
DE 19851415A1
· 2000
[cited by applicant]
EP 1071776A2
· 2001
[cited by applicant]
EP 1358349A2
· 2003
[cited by applicant]
EP 1394274A2
· 2004
[cited by applicant]
EP 1572987A2
· 2005
[cited by applicant]
EP 1578367A2
· 2005
[cited by applicant]
EP 1578996A2
· 2005
[cited by applicant]
EP 1888627A2
· 2008
[cited by applicant]
EP 2103685A1
· 2008
[cited by applicant]
EP 2096169B1
· 2008
[cited by applicant]
EP 2021499A2
· 2009
[cited by applicant]
EP 2126135A2
· 2009
[cited by applicant]
EP 2132225A1
· 2009
[cited by applicant]
EP 2435557A1
· 2010
[cited by applicant]
EP 2191018A2
· 2010
[cited by applicant]
EP 2191840A1
· 2010
[cited by applicant]
EP 2612911B1
· 2011
[cited by applicant]
EP 2388336A1
· 2011
[cited by applicant]
EP 2407488A2
· 2012
[cited by applicant]
EP 2421563A1
· 2012
[cited by applicant]
EP 2432881A2
· 2012
[cited by applicant]
EP 2478101A1
· 2012
[cited by applicant]
EP 2572000A2
· 2013
[cited by applicant]
EP 2638163A1
· 2013
[cited by applicant]
EP 2655621A1
· 2013
[cited by applicant]
EP 2675903A1
· 2013
[cited by applicant]
EP 2852671A2
· 2015
[cited by applicant]
EP 2931914A1
· 2015
[cited by applicant]
EP 3060237A1
· 2016
[cited by applicant]
EP 3452101A2
· 2017
[cited by applicant]
EP 3194623A1
· 2017
[cited by applicant]
EP 2643459B1
· 2017
[cited by applicant]
EP 3334755A1
· 2018
[cited by applicant]
EP 3385373A1
· 2018
[cited by applicant]
WO WO9954460A2
· 1999
[cited by applicant]
WO WO2000069450A1
· 2000
[cited by applicant]
WO WO2001094629A2
· 2001
[cited by applicant]
WO WO2004073657A2
· 2004
[cited by applicant]
WO WO2005052164A1
· 2005
[cited by applicant]
WO WO2006123930A2
· 2006
[cited by applicant]
WO WO2007058527A2
· 2007
[cited by applicant]
WO WO2007078599A2
· 2007
[cited by applicant]
WO WO2008051854A2
· 2008
[cited by applicant]
WO WO2008081435A2
· 2008
[cited by applicant]
WO WO2009028945A2
· 2009
[cited by applicant]
WO WO2009061442A1
· 2009
[cited by applicant]
WO WO2010030003A1
· 2009
[cited by applicant]
WO WO2010104357A2
· 2010
[cited by applicant]
WO WO2010123501A1
· 2010
[cited by applicant]
WO WO2010135329A2
· 2010
[cited by applicant]
WO WO2010138263A2
· 2010
[cited by applicant]
WO WO2011017910A1
· 2011
[cited by applicant]
WO WO2011034421A1
· 2011
[cited by applicant]
WO WO2011145615A1
· 2011
[cited by applicant]
WO WO2011144718A2
· 2011
[cited by applicant]
WO WO2012014207A2
· 2012
[cited by applicant]
WO WO2012065143A1
· 2012
[cited by applicant]
WO WO2012071549A2
· 2012
[cited by applicant]
WO WO2012087983A1
· 2012
[cited by applicant]
WO WO2012120026A1
· 2012
[cited by applicant]
WO WO2012136841A1
· 2012
[cited by applicant]
WO WO2013177133A2
· 2013
[cited by applicant]
WO WO2014053082A1
· 2014
[cited by applicant]
WO WO2014152607A2
· 2014
[cited by applicant]
WO WO2015077498A1
· 2014
[cited by applicant]
WO WO2014191391A1
· 2014
[cited by applicant]
WO WO2016171448A1
· 2016
[cited by applicant]
WO WO2016170348A2
· 2016
[cited by applicant]
WO WO2017026776A1
· 2017
[cited by applicant]
WO WO2017173354A2
· 2017
[cited by applicant]
WO WO2017180587A2
· 2017
[cited by applicant]
WO WO2018041959A1
· 2018
[cited by applicant]
WO WO2019053012A1
· 2018
[cited by applicant]
WO WO2018204764A1
· 2018
[cited by applicant]
WO WO2019023680A1
· 2019
[cited by examiner]
WO WO2019094778A1
· 2019
[cited by applicant]
WO WO2019099552A1
· 2019
[cited by examiner]
WO WO2020012164A1
· 2020
[cited by applicant]
WO WO2020069339A1
· 2020
[cited by applicant]
WO WO2020069373A1
· 2020
[cited by applicant]
WO WO2021183825A1
· 2021
[cited by applicant]
WO WO2021183946A1
· 2021
[cited by applicant]
WO WO2022232327A2
· 2022
[cited by applicant]
WO WO2023004367A2
· 2023
[cited by applicant]
WO WO2023196851A1
· 2023
[cited by applicant]
Roney IJ, Rudner AD, Couture JF, Karn M. Improvement of the reverse tetracycline transactivator by single amino acid substitutions that reduce leaky target gene expression to undetectable levels. Sci Rep. 2016;6:27697. …
[cited by examiner]
Matsui, R., Tanabe, Y., & Watanabe, D. (2012). Avian adeno-associated virus vector efficiently transduces neurons in the embryonic and post-embryonic chicken brain. PLoS One, 7(11), e48730. (Year: 2012).
[cited by examiner]
Choi, Jun-Hyeok, et al. “Optimization of AAV expression cassettes to improve packaging capacity and transgene expression in neurons.” Molecular brain 7 (2014): 1-10. (Year: 2014).
[cited by examiner]
Fan, Xiujun, et al. “Transient, inducible, placenta-specific gene expression in mice.” Endocrinology 153.11 (2012): 5637-5644. (Year : 2012).
[cited by examiner]
Kim, C. H., Oh, Y., & Lee, T. H. (1997). Codon optimization for high-level expression of human erythropoietin (EPO) in mammalian cells. Gene, 199(1-2), 293-301. (Year: 1997).
[cited by examiner]
International Search Report and Written Opinion for Application No. PCT/US2019/053545, mailed Dec. 19, 2019.
[cited by applicant]
International Preliminary Report on Patentability for Application No. PCT/US2019/053545, mailed Apr. 8, 2021.
[cited by applicant]
International Search Report and Written Opinion for Application No. PCT/US2019/053492, mailed Feb. 2, 2020.
[cited by applicant]
International Preliminary Report on Patentability for Application No. PCT/US2019/053492, mailed Apr. 8, 2021.
[cited by applicant]
No Author Listed, Tet-On® 3G Inducible Expression System. Clontech Laboratories, Inc. 8 pages.
[cited by applicant]
Agha-Mohammadi et al., Second-generation tetracycline-regulatable promoter: repositioned tet operator elements optimize transactivator synergy while shorter minimal promoter offers tight basal leakiness. J Gene Med. Jul…
[cited by applicant]
Aida et al., Cloning-free CRISPR/Cas system facilitates functional cassette knock-in in mice. Genome Biol. Apr. 29, 2015;16(1):87. doi: 10.1186/s13059-015-0653-x.
[cited by applicant]
Alaei et al., An improved reprogrammable mouse model harbouring the reverse tetracycline-controlled transcriptional transactivator 3. Stem Cell Res. Jul. 2016;17(1):49-53. doi: 10.1016/j.scr.2016.05.008. Epub May 25, 20…
[cited by applicant]
Azte et al., Selecting the optimal Tet-On system for doxycycline-inducible gene expression in transiently transfected and stably transduced mammalian cells. Biotechnol J. Jan. 2016;11(1):71-9. doi: 10.1002/biot.20150023…
[cited by applicant]
Bar-Nur et al., Small molecules facilitate rapid and synchronous iPSC generation. Nat Methods. Nov. 2014;11(11):1170-6. doi: 10.1038/nmeth.3142. Epub Sep. 24, 2014.
[cited by applicant]
Baron et al., Tet repressor-based system for regulated gene expression in eukaryotic cells: principles and advances. Methods Enzymol. 2000;327:401-21. doi: 10.1016/s0076-6879(00)27292-3.
[cited by applicant]
Bussian et al., Clearance of senescent glial cells prevents tau-dependent pathology and cognitive decline. Nature. Oct. 2018;562(7728):578-582. doi: 10.1038/s41586-018-0543-y. Epub Sep. 19, 2018.
[cited by applicant]
Carey et al., Reprogramming of murine and human somatic cells using a single polycistronic vector. Proc Natl Acad Sci U S A. Jan. 6, 2009;106(1):157-62. doi: 10.1073/pnas.0811426106. Epub Dec. 24, 2008.
[cited by applicant]
Cho et al., Generation of transgenic mice. Curr Protoc Cell Biol. Mar. 2009;Chapter 19:Unit 19.11. doi: 10.1002/0471143030.cb1911s42.
[cited by applicant]
Danke et al., Adjusting transgene expression levels in lymphocytes with a set of inducible promoters. J Gene Med. Jun. 2010;12(6):501-15. doi: 10.1002/jgm.1461.
[cited by applicant]
Das et al., Selecting the optimal Tet-On system for doxycycline-inducible gene expression in transiently transfected and stably transduced mammalian cells. Biotechnol J. Jan. 2016;11(1):71-9. doi: 10.1002/biot.201500236…
[cited by applicant]
Doench et al., Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nat Biotechnol. Feb. 2016;34(2):184-191. doi: 10.1038/nbt.3437. Epub Jan. 18, 2016.
[cited by applicant]
Dong et al., Poly(glycoamidoamine) Brushes Formulated Nanomaterials for Systemic siRNA and mRNA Delivery in Vivo. Nano Lett. Feb. 10, 2016;16(2):842-8. doi: 10.1021/acs.nanolett.5b02428. Epub Jan. 13, 2016.
[cited by applicant]
Gossen et al., Transcriptional activation by tetracyclines in mammalian cells. Science. Jun. 23, 1995;268(5218):1766-9. doi: 10.1126/science.7792603.
[cited by applicant]
Hsu et al., Development and applications of CRISPR-Cas9 for genome engineering. Cell. Jun. 5, 2014;157(6):1262-1278. doi: 10.1016/j.cell.2014.05.010.
[cited by applicant]
Jiang et al., Tetracycline-regulated gene expression mediated by a novel chimeric repressor that recruits histone deacetylases in mammalian cells. J Biol Chem. Nov. 30, 2001;276(48):45168-74.
[cited by applicant]
Lozano-Torres et al., An OFF-ON Two-Photon Fluorescent Probe for Tracking Cell Senescence in Vivo. J Am Chem Soc. Jul. 5, 2017;139(26):8808-8811. doi: 10.1021/jacs.7b04985. Epub Jun. 23, 2017.
[cited by applicant]
Manukyan et al., Epigenome rejuvenation: HP1β mobility as a measure of pluripotent and senescent chromatin ground states. Sci Rep. Apr. 25, 2014;4:4789. doi: 10.1038/srep04789.
[cited by applicant]
Moreira et al., Assessing Executive Dysfunction in Neurodegenerative Disorders: A Critical Review of Brief Neuropsychological Tools. Front Aging Neurosci. Nov. 9, 2017;9:369. doi: 10.3389/fnagi.2017.00369. eCollection 2…
[cited by applicant]
Nehlin et al., The Werner syndrome. A model for the study of human aging. Ann N Y Acad Sci. Jun. 2000;908:167-79. doi: 10.1111/j.1749-6632.2000.tb06645.x.
[cited by applicant]
Ocampo et al., In Vivo Amelioration of Age-Associated Hallmarks by Partial Reprogramming. Cell. Dec. 15, 2016;167(7):1719-1733.e12. doi: 10.1016/j.cell.2016.11.052.
[cited by applicant]
O'Connor et al., Genetic medicines: treatment strategies for hereditary disorders. Nat Rev Genet. Apr. 2006;7(4):261-76. doi: 10.1038/nrg1829.
[cited by applicant]
Ramamoorth et al., Non viral vectors in gene therapy—an overview. J Clin Diagn Res. Jan. 2015;9(1):GE01-6. doi: 10.7860/JCDR/2015/10443.5394. Epub Jan. 1, 2015.
[cited by applicant]
Roney et al., Improvement of the reverse tetracycline transactivator by single amino acid substitutions that reduce leaky target gene expression to undetectable levels. Sci Rep. Jun. 21, 2016;6:27697. doi: 10.1038/srep2…
[cited by applicant]
Sander et al., CRISPR-Cas systems for editing, regulating and targeting genomes. Nat Biotechnol. Apr. 2014;32(4):347-55. doi: 10.1038/nbt.2842. Epub Mar. 2, 2014.
[cited by applicant]
Sarin et al., Inhibition of acquired immunodeficiency syndrome virus by oligodeoxynucleoside methylphosphonates. Proc Natl Acad Sci USA. Oct. 1988;85(20):7448-51. Doi 10.1073/pnas.85.20.7448.
[cited by applicant]
Senis et al, AAV vector-mediated in vivo reprogramming into pluripotency. Nat Commun. Jul. 9, 2018;9(1):2651. doi: 10.1038/s41467-018-05059-x.
[cited by applicant]
Smalley, First AAV gene therapy poised for landmark approval. Nat Biotechnol. Nov. 9, 2017;35(11):998-999. doi: 10.1038/nbt1117-998.
[cited by applicant]
Tammam et al., Nuclear delivery of recombinant OCT4 by chitosan nanoparticles for transgene-free generation of protein-induced pluripotent stem cells. Oncotarget. Jun. 21, 2016;7(25):37728-37739. doi: 10.18632/oncotarge…
[cited by applicant]
Tyner et al., p53 mutant mice that display early ageing-associated phenotypes. Nature. Jan. 3, 2002;415(6867):45-53. doi: 10.1038/415045a.
[cited by applicant]
Wang et al., Spatiotemporal control of gene expression by a light-switchable transgene system. Nat Methods. Feb. 12, 2012;9(3):266-9. doi: 10.1038/nmeth.1892.
[cited by applicant]
Wright et al., Identification of factors that contribute to recombinant AAV2 particle aggregation and methods to prevent its occurrence during vector purification and formulation. Mol Ther. Jul. 2005;12(1):171-8. doi: 1…
[cited by applicant]
Yilmazer et al., In vivo cell reprogramming towards pluripotency by virus-free overexpression of defined factors. PLoS One. 2013;8(1):e54754. doi: 10.1371/journal.pone.0054754. Epub Jan. 23, 2013.
[cited by applicant]
Zahid et al., Protein transduction domains: applications for molecular medicine. Curr Gene Ther. Oct. 2012;12(5):374-80. doi: 10.2174/156652312802762527.
[cited by applicant]
Zhao et al., A coumermycin/novobiocin-regulated gene expression system. Hum Gene Ther. Nov. 20, 2003;14(17):1619-29. doi: 10.1089/104303403322542266.
[cited by applicant]
Zhou et al., Generation of induced pluripotent stem cells using recombinant proteins. Cell Stem Cell. May 8, 2009;4(5):381-4. doi: 10.1016/j.stem.2009.04.005. Epub Apr. 23, 2009.
[cited by applicant]
Zhou et al., Optimization of the Tet-On system for regulated gene expression through viral evolution. Gene Ther. Oct. 2006;13(19):1382-90. doi: 10.1038/sj.gt.3302780. Epub May 25, 2006.
[cited by applicant]
International Search Report and Written Opinion for Application No. PCT/US2023/065374, mailed Jun. 6, 2023.
[cited by applicant]
Chtarto et al., A regulatable AAV vector mediating GDNF biological effects at clinically-approved sub-antimicrobial doxycycline doses. Mol Ther Methods Clin Dev. Mar. 30, 2016:5:16027. doi: 10.1038/mtm.2016.27. eCollect…
[cited by applicant]
Das et al., Tet-On Systems for Doxycycline-inducible Gene Expression. Curr Gene Ther. 2016;16(3):156-67. doi: 10.2174/1566523216666160524144041.
[cited by applicant]
Gill et al., Multi-omic rejuvenation of human cells by maturation phase transient reprogramming. Elife. Apr. 8, 2022:11:e71624. doi: 10.7554/eLife.71624.
[cited by applicant]
Gossen et al., Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. Jun. 15, 1992;89(12):5547-51. doi: 10.1073/pnas.89.12.5547.
[cited by applicant]
Heinz et al., Retroviral and transposon-based tet-regulated all-in-one vectors with reduced background expression and improved dynamic range. Hum Gene Ther. Feb. 2011;22(2):166-76. doi: 10.1089/hum.2010.099. Epub Dec. 1…
[cited by applicant]
Hishida et al., In vivo partial cellular reprogramming enhances liver plasticity and regeneration. Cell Rep. Apr. 26, 2022;39(4):110730. doi: 10.1016/j.celrep.2022.110730.
[cited by applicant]
Hosoda et al., Development of a tightly-regulated tetracycline-dependent transcriptional activator and repressor co-expression system for the strong induction of transgene expression. Cytotechnology. May 2011;63(3):211-…
[cited by applicant]
Karg et al., Sustained vision recovery by OSK gene therapy in a mouse model of glaucoma. Cell Reprogram. Dec. 2023;25(6):288-299. doi: 10.1089/cell.2023.0074. Epub Dec. 7, 2023.
[cited by applicant]
Loew et al., Improved Tet-responsive promoters with minimized background expression. BMC Biotechnol. Nov. 24, 2010:10:81. doi: 10.1186/1472-6750-10-81.
[cited by applicant]
Lu et al., Reprogramming to recover youthful epigenetic information and restore vision. Nature. Dec. 2020;588(7836):124-129. doi: 10.1038/s41586-020-2975-4. Epub Dec. 2, 2020. with Supplementary Information. 35 pages.
[cited by applicant]
Michalon et al., Inducible and neuron-specific gene expression in the adult mouse brain with the rtTA2S-M2 system. Genesis. Dec. 2005;43(4):205-12. doi: 10.1002/gene.20175.
[cited by applicant]
No Author Listed, Tet-On® 3G Inducible Expression Systems User Manual. Clonetech Laboratories, Inc. 2014. 24 pages. Published online at takarabio.com.
[cited by applicant]
No Author Listed, Tet-One technology overview. Takara Bio USA, Inc. Accessed at: https://www.takarabio.com/learning-centers/gene-function/inducible-systems/tet-inducible-systems/tet-one-technology-overview. Last accesse…
[cited by applicant]
Pico et al., Comparative analysis of mouse strains for in vivo reprogramming. bioRxiv. Mar. 8, 2024. 32 pages. https://doi.org/10.1101/2024.03.08.584074.
[cited by applicant]
Randolph et al., An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives. Sci Rep. May 8, 2017;7(1):1549. doi: 10.1038/s41598-017-01684-6.
[cited by applicant]
Sheng et al., Generation and characterization of a Tet-On (rtTA-M2) transgenic rat. BMC Dev Biol. Feb. 16, 2010:10:17. doi: 10.1186/1471-213X-10-17.
[cited by applicant]
Takahashi et al., Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. Nov. 30, 2007;131(5):861-72. doi: 10.1016/j.cell.2007.11.019.
[cited by applicant]
Takahashi et al., Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. Aug. 25, 2006;126(4):663-76. doi: 10.1016/j.cell.2006.07.024. Epub Aug. 10, 2006.
[cited by applicant]
Zhang et al., Reducing Background Expression of Target Gene with tTS/rtTA System in Cell Model. Journal of Sun Yat-Sen University (Medical Sciences). Jul. 30, 2006; 27(4): 361-364.
[cited by applicant]
Zhu et al., Silencing and un-silencing of tetracycline-controlled genes in neurons. PLoS One. Jun. 20, 2007;2(6):e533. doi: 10.1371/journal.pone.0000533.
[cited by applicant]
Lamartina et al., Construction of an rtTA2(s)-m2/tts(kid)-based transcription regulatory switch that displays No. basal activity, good inducibility, and high responsiveness to doxycycline in mice and non-human primates.…
[cited by applicant]
Uchida et al., Tight regulation of transgene expression by tetracycline-dependent activator and repressor in brain. Genes Brain Behav. Feb. 2006;5(1):96-106. doi: 10.1111/j.1601-183X.2005.00139.x.
[cited by applicant]
Urlinger et al., Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc Natl Acad Sci U S A. Jul. 5, 2000;97(14):7963-8. doi: 10.10…
[cited by applicant]
Yao et al., Systemic and localized reversible regulation of transgene expression by tetracycline with tetR-mediated transcription repression switch. J Surg Res. Apr. 2007;138(2):267-74. doi: 10.1016/j.jss.2006.05.007. E…
[cited by applicant]
U.S. Appl. No. 18/854,506, filed Oct. 4, 2024, Sinclair et al.
[cited by applicant]
International Preliminary Report on Patentability for Application No. PCT/US2023/065374, mailed Oct. 17, 2024.
[cited by applicant]
[No Author Listed],
[cited by applicant]
[No Author Listed], Optic Nerve Crush. EYECRO. 2024. Accessed at: https://eyecro.com/models/optic-nerve-crush/. [last accessed: Sep. 18, 2024].
[cited by applicant]
Bareyre et al., In vivo imaging reveals a phase-specific role of STAT3 during central and peripheral nervous system axon regeneration. Proc Natl Acad Sci USA. Apr. 12, 2011;108(15):6282-7. doi: 10.1073/pnas.1015239108. …
[cited by applicant]
Bekris et al., The Genetics of Parkinson's Disease. J Geriatr Psychiatry Neurol. Dec. 2010;23(4):228-42. doi: 10.1177/0891988710383572. Epub Oct. 11, 2010.
[cited by applicant]
Brennan et al., Ocular Salvage and Vision Preservation Using a Topotecan-Based Regimen for Advanced Intraocular Retinoblastoma. J Clin Oncol. Jan. 2017;35(1):72-77. doi: 10.1200/JCO.2016.69.2996. Epub Oct. 31, 2016.
[cited by applicant]
Cameron et al., Optic Nerve Crush in Mice to Study Retinal Ganglion Cell Survival and Regeneration. Bio Protoc. Mar. 20, 2020;10(6):e3559. doi: 10.21769/BioProtoc.3559.
[cited by applicant]
Chan et al., Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Nat Neurosci. Author manuscript; available in PMC: Dec. 26, 2017. Published in final edited form as: Na…
[cited by applicant]
Chtarto et al., Tetracycline-inducible transgene expression mediated by a single AAV vector. Gene Ther. Jan. 2003;10(1):84-94. doi: 10.1038/sj.gt.3301838.
[cited by applicant]
Ellis et al., A survey of ex vivo/in vitro transduction efficiency of mammalian primary cells and cell lines with Nine natural adeno-associated virus (AAV1-9) and one engineered adeno-associated virus serotype. Virol J.…
[cited by applicant]
Fan et al., Transient, Inducible, Placenta-Specific Gene Expression in Mice. Endocrinology. Nov. 2012;153(11):5637-44. doi: 10.1210/en.2012-1556. Epub Sep. 25, 2012.
[cited by applicant]
Fardi et al., Epigenetic mechanisms as a new approach in cancer treatment: An updated review. Genes Dis. Jun. 18, 2018;5(4):304-311. doi: 10.1016/j.gendis.2018.06.003. eCollection Dec. 2018.
[cited by applicant]
Gjoneska et al., Conserved epigenomic signals in mice and humans reveal immune basis of Alzheimer's disease. Nature. Feb. 19, 2015;518(7539):365-9. doi: 10.1038/nature14252.
[cited by applicant]
Goertsen et al., AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset. Nat Neurosci. Jan. 2022;25(1):106-115. doi: 10.1038/s41593-021-00…
[cited by applicant]
Hager et al., An internal polyadenylation signal substantially increases expression levels of lentivirus-delivered transgenes but has the potential to reduce viral titer in a promoter-dependent manner. Hum Gene Ther. Au…
[cited by applicant]
Khalilpour et al., Ischemic optic neuropathy as a model of neurodegenerative disorder: A review of pathogenic mechanism of axonal degeneration and the role of neuroprotection. J Neurol Sci. Apr. 15, 2017:375:430-441. do…
[cited by applicant]
Krolak et al., A High-Efficiency AAV for Endothelial Cell Transduction Throughout the Central Nervous System. Nat Cardiovasc Res. Author manuscript; available in PMC: Oct. 13, 2022. Published in final edited form as: Na…
[cited by applicant]
Lambert et al., Towards a Microbead Occlusion Model of Glaucoma for a Non-Human Primate. Sci Rep. Aug. 9, 2019;9(1):11572. doi: 10.1038/s41598-019-48054-y.
[cited by applicant]
Li et al., Epigenetics and Common Ophthalmic Diseases. Yale J Biol Med. Dec. 23, 2016;89(4):597-600. eCollection Dec. 2016.
[cited by applicant]
Li et al., Production of Lentiviral Vectors for Transducing Cells from the Central Nervous System. J Vis Exp. May 24, 2012:(63):e4031. doi: 10.3791/4031.
[cited by applicant]
Mahmoudi et al., Old fibroblasts secrete inflammatory cytokines that drive variability in reprogramming efficiency and may affect wound healing between old individuals. Biorxiv. Oct. 19, 2018. doi: https://doi.org/10.11…
[cited by applicant]
Mathiesen et al., CNS Transduction Benefits of AAV-PHP.eB over AAV9 Are Dependent on Administration Route and Mouse Strain. Mol Ther Methods Clin Dev. Oct. 20, 2020:19:447-458. doi: 10.1016/j.omtm.2020.10.011. eCollecti…
[cited by applicant]
Mertens et al., Directly Reprogrammed Human Neurons Retain Aging-Associated Transcriptomic Signatures and Reveal Age-Related Nucleocytoplasmic Defects. Cell Stem Cell. Dec. 3, 2015;17(6):705-718. doi: 10.1016/j.stem.201…
[cited by applicant]
Nakagawa et al., Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat Biotechnol. Jan. 2008;26(1):101-6. doi: 10.1038/nbt1374. Epub Nov. 30, 2007.
[cited by applicant]
Olova et al., Partial reprogramming induces a steady decline in epigenetic age before loss of somatic identity. Aging Cell. Feb. 2019;18(1):e12877. doi: 10.1111/acel.12877. Epub Nov. 18, 2018.
[cited by applicant]
Reichmuth et al., mRNA vaccine delivery using lipid nanoparticles. Ther Deliv. 2016;7(5):319-34. doi: 10.4155/tde-2016-0006.
[cited by applicant]
Shannon, A Mathematical Theory of Communication. The Bell System Technical Journal. 1948; 1: 379-423. https://doi.org/10.1002/j.1538-7305.1948.tb01338.x.
[cited by applicant]
Wang et al., 2A self-cleaving peptide-based multi-gene expression system in the silkworm Bombyx mori. Sci Rep. Nov. 5, 2015:5:16273. doi: 10.1038/srep16273.
[cited by applicant]
Abad et al., Reprogramming in vivo produces teratomas and iPS cells with totipotency features. Nature. Oct. 17, 2013;502(7471):340-5. doi: 10.1038/nature12586. Epub Sep. 11, 2013.
[cited by applicant]
Hansson et al., Highly coordinated proteome dynamics during reprogramming of somatic cells to pluripotency. Cell Rep. Dec. 27, 2012;2(6):1579-92. doi: 10.1016/j.celrep.2012.10.014.
[cited by applicant]
Karg et al., Partial epigenetic reprogramming of RPE cells reverses age and rejuvenates mitochondrial metabolism. Investigative Ophthalmology & Visual Science. Jun. 2023; 64(8): 2966. ARVO Annual Meeting Abstract.
[cited by applicant]
Ksander et al., Epigenetic reprogramming-A novel gene therapy that restores vision loss in a nonhuman primate model of NAION. Investigative Ophthalmology & Visual Science. Jun. 2023; 64(8): 474. ARVO Annual Meeting Abst…
[cited by applicant]
Kurian et al., Conversion of human fibroblasts to angioblast-like progenitor cells. Nat Methods. Jan. 2013;10(1):77-83. doi: 10.1038/nmeth.2255. Epub Dec. 2, 2012.
[cited by applicant]
Ohnishi et al., Premature termination of reprogramming in vivo leads to cancer development through altered epigenetic regulation. Cell. Feb. 13, 2014;156(4):663-77. doi: 10.1016/j.cell.2014.01.005.
[cited by applicant]
Polo et al., A molecular roadmap of reprogramming somatic cells into iPS cells. Cell. Dec. 21, 2012;151(7):1617-32. doi: 10.1016/j.cell.2012.11.039.
[cited by applicant]
Roux et al., Diverse partial reprogramming strategies restore youthful gene expression and transiently suppress cell identity. Cell Syst. Jul. 20, 2022;13(7):574-587.e11. doi: 10.1016/j.cels.2022.05.002. Epub Jun. 10, 2…
[cited by applicant]
Roux et al., Partial reprogramming restores youthful gene expression through transient suppression of cell identity. bioRxiv. May 23, 2021. DOI: 10.1101/2021.05.21.444556.
[cited by applicant]
Sahu et al., Targeted partial reprogramming of age-associated cell states improves markers of health in mouse models of aging. Sci Transl Med. Sep. 11, 2024;16(764):eadg1777. doi: 10.1126/scitranslmed.adg1777. Epub Sep.…
[cited by applicant]
Soldner et al., Parkinson's disease patient-derived induced pluripotent stem cells free of viral reprogramming factors. Cell. Mar. 6, 2009;136(5):964-77. doi: 10.1016/j.cell.2009.02.013.
[cited by applicant]
Sun et al., Tetracycline-inducible expression systems: new strategies and practices in the transgenic mouse modeling. Acta Biochim Biophys Sin (Shanghai). Apr. 2007;39(4):235-46. doi: 10.1111/j.1745-7270.2007.00258.x.
[cited by applicant]
Thier et al., Direct conversion of fibroblasts into stably expandable neural stem cells. Cell Stem Cell. Apr. 6, 2012;10(4):473-9. doi: 10.1016/j.stem.2012.03.003. Epub Mar. 22, 2012.
[cited by applicant]
Vivien et al., Non-viral expression of mouse Oct4, Sox2, and Klf4 transcription factors efficiently reprograms tadpole muscle fibers in vivo. J Biol Chem. Mar. 2, 2012;287(10):7427-35. doi: 10.1074/jbc.M111.324368. Epub…
[cited by applicant]
U.S. Appl. No. 18/318,566, filed May 16, 2023, Sinclair et al.
[cited by applicant]
U.S. Appl. No. 18/583,191, filed Feb. 21, 2024, Sinclair et al.
[cited by applicant]
U.S. Appl. No. 18/583,147, filed Feb. 21, 2024, Sinclair et al.
[cited by applicant]