Granted Patent
B2
US 12,350,284 · App. 17/051,578 · Granted Jul 8, 2025
BCL11A microRNAs for treating hemoglobinopathies
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Abstract
Embodiments herein provide specially designed synthetic BCL11A-targeting microRNAs for RNA polymerase II expression, and methods of use to treat hemoglobinopathies, such as sickle cell disease or thalassemia, by increasing the expression levels of fetal hemoglobin levels.
Claims (17)
1. A synthetic BCL11A microRNA comprising;
a) a first BCL11A segment,
b) a loop segment derived from miRNA144; and
c) a second BCL11A segment arranged in tandem in a 5′ to 3′ direction,
wherein the loop segment is between and directly linked to the first and second BCL11A segments;
wherein the second BCL11A segment is complementary to the first BCL11A segment so that the first and second BCL11A segments base pair to form a hairpin loop with the loop segment forming the loop portion of the hairpin loop thus formed; and
wherein the first BCL11A segment starts with a -GCGC- at the 5′ end and the second BCL11A segment ends with a -GCGC- at the 3′ end,
wherein the loop segment consists of the sequence of AGTTTGCGATGAGACAC (SEQ ID NO: 68).
2. The synthetic BCL11A microRNA of claim 1 , wherein the first and second BCL11A segments are 18 to 25 nucleotides long.
3. The synthetic BCL11A microRNA of claim 1 , wherein the first BCL11A segment contains a sequence from a BCL11A mRNA sequence.
4. The synthetic BCL11A microRNA of claim 1 , wherein the first BCL11A segment comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 57, 58, 59, 60, 61, 62, 63, 64, and 65.
5. The synthetic BCL11A microRNA of claim 1 , wherein the first BCL11A segment comprises the nucleotide sequence set forth in SEQ ID NO: 61.
6. The synthetic BCL11A microRNA of claim 1 , wherein the microRNA comprising a nucleotide sequence consisting of SEQ ID NO: 98.
7. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 98.
8. A vector comprising the isolated nucleic acid molecule of claim 7 .
9. A isolated host cell comprising the vector of claim 8 .
10. The isolated host cell of claim 9 , wherein the cell is a hematopoietic stem cell or progenitor cell.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Mar 6, 2025
From: WILLIAMS, DAVID A.; GUDA, SWAROOPA
To: CHILDREN'S MEDICAL CENTER CORPORATION, THE
Reel/Frame 070423/0918 →
Continuity (2)
Provisional Application
62665691
· May 2, 2018
References Cited (206)
US 5460964A
· McGlave et al.
· 1995
[cited by applicant]
US 5635387A
· Fei et al.
· 1997
[cited by applicant]
US 5677136A
· Simmons et al.
· 1997
[cited by applicant]
US 5716827A
· Tsukamoto et al.
· 1998
[cited by applicant]
US 5750397A
· Tsukamoto et al.
· 1998
[cited by applicant]
US 5759793A
· Schwartz et al.
· 1998
[cited by applicant]
US 5928638A
· Uchida et al.
· 1999
[cited by applicant]
US 6682907B1
· Charneau et al.
· 2004
[cited by applicant]
US 8383604B2
· Orkin et al.
· 2013
[cited by applicant]
US 9228185B2
· Orkin et al.
· 2016
[cited by applicant]
US 9822355B2
· Orkin et al.
· 2017
[cited by applicant]
US 9885041B2
· Orkin et al.
· 2018
[cited by applicant]
US 20040259247A1
· Tuschl et al.
· 2004
[cited by applicant]
US 20080051431A1
· Verhelle et al.
· 2008
[cited by applicant]
US 20110294114A1
· Van Der Loo et al.
· 2011
[cited by applicant]
US 20150232882A1
· Zheng et al.
· 2015
[cited by applicant]
EP 1752536A1
· 2007
[cited by applicant]
EP 2334794B1
· 2016
[cited by applicant]
JP 2006507841A
· 2006
[cited by applicant]
WO 2004054512A2
· 2004
[cited by applicant]
WO 2009007685A2
· 2009
[cited by applicant]
WO 2010030963A2
· 2010
[cited by applicant]
WO 2011072086A1
· 2011
[cited by applicant]
WO 2011133889A2
· 2011
[cited by applicant]
WO 2012073047A2
· 2012
[cited by applicant]
WO 2012079046A2
· 2012
[cited by applicant]
WO 2013049615A1
· 2013
[cited by applicant]
WO 2013126794A1
· 2013
[cited by applicant]
WO 2013176772A1
· 2013
[cited by applicant]
WO 2014085593A1
· 2014
[cited by applicant]
WO 2014093965A1
· 2014
[cited by applicant]
WO 2015065964A1
· 2015
[cited by applicant]
WO 2015148863A2
· 2015
[cited by applicant]
WO 2015164739A1
· 2015
[cited by applicant]
WO 2015164750A2
· 2015
[cited by applicant]
WO 2015164759A2
· 2015
[cited by applicant]
WO 2015183667A1
· 2015
[cited by applicant]
WO 2016094304A2
· 2016
[cited by applicant]
WO 2016182893A1
· 2016
[cited by applicant]
WO 2016183448A1
· 2016
[cited by applicant]
WO 2017040529A1
· 2017
[cited by applicant]
WO 2334794B8
· 2017
[cited by applicant]
WO 2017115268A1
· 2017
[cited by applicant]
WO 2017139576A1
· 2017
[cited by applicant]
WO 2017173092A1
· 2017
[cited by applicant]
WO 2017182881A2
· 2017
[cited by applicant]
WO 2018218135A1
· 2018
[cited by applicant]
AEB92750, Croce et al. (WO 2005078139) (Year: 2005).
[cited by examiner]
Akinsheye et al., “Fetal hemoglobin in sickle cell anemia.” Blood 118(1):19-27 (2011).
[cited by applicant]
Amaya et al., “Mi2β-mediated silencing of the fetal γ-globin gene in adult erythroid cells.” Blood 121(17):3493-501 (2013).
[cited by applicant]
Amendah et al., “Sickle cell disease-related pediatric medical expenditures in the U.S.” American Journal of Preventive Medicine 38(4 Suppl):S550-S556 (2010).
[cited by applicant]
Atweh et al., “Pharmacological induction of fetal hemoglobin in sickle cell disease and beta-thalassemia.” Seminars in Hematology 38(4):367-73 (2001).
[cited by applicant]
Bauer et al., “An Erythroid Enhancer of BCL11A Subject to Genetic Variation Determines Fetal Hemoglobin Level.” Science 342(6155):253-257 (2013).
[cited by applicant]
Bauer et al., “HbF-Associated Genetic Variation Marks an Erythroid Regulatory Element Essential for BCL11A Transcription and Subsequent Stage-Specific Globin Expression.” Blood 120:828 (2012).
[cited by applicant]
Bauer et al., “Hemoglobin switching's surprise: the versatile transcription factor BCL11A is a master repressor of fetal hemoglobin” Current Opinion in Genetics & Development 33:62-70 (2015).
[cited by applicant]
Bauer et al., “Reawakening fetal hemoglobin: prospects for new therapies for the B-globin disorders.” Blood 120 (15):2945-2953 (2012).
[cited by applicant]
Bauer et al., “Supplementary Material: An Erythroid Enhancer of BCL11A Subject to Genetic Variation Determines Fetal Hemoglobin Level.” Science 342(6155):253-257 (2013).
[cited by applicant]
Bjurström et al. “Reactivating fetal hemoglobin expression in human adult erythroblasts through BCL11A knockdown using targeted endonucleases.” Molecular Therapy—Nucleic Acids 5:e351 (2016).
[cited by applicant]
Boettcher et al., “Choosing the right tool for the job: RNAi, Talen, or CRISPR.” Molecular Cell 58(4):575-585 (2015).
[cited by applicant]
Bohmer et al., “Identification of fetal nucleated red cells in co-cultures from fetal and adult peripheral blood: differential effects of serum on fetal and adult erythropoiesis.” Prenatal Diagnosis 19(7):628-636 (1999).
[cited by applicant]
Bunn “Reversing ontogeny.” New Engl. J. Med. 328(2):129-131 (1993).
[cited by applicant]
Canver et al., “BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis.” Nature 527 (7577):192-197 (2015).
[cited by applicant]
Cao et al., “Recent advances in B-thalassemias.” Pediatric Reports 3(2):65-71 (2011).
[cited by applicant]
Chabchoub et al., “The facial dysmorphy in the newly recognised microdeletion 2p15-p16.1 refined to a 570 kb region in 2p15.” Journal of Medical Genetics 45(3):189-192 (2008).
[cited by applicant]
Coleman et al., “Sickle cell anemia: targeting the role of fetal hemoglobin in therapy.” Clinical Pediatrics 46 (5):386-391 (2007).
[cited by applicant]
Cox et al., “Therapeutic genome editing: prospects and challenges” Nature Medicine 21(2):121-131 (2015).
[cited by applicant]
Dixit et al., “Hydroxyurea in thalassemia intermedia—a promising therapy.” Annals of Hematology 84(7):441-446 (2005).
[cited by applicant]
Doench et al., “Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation.” Nature Biotechnology 32(12):1262-1267 (2014).
[cited by applicant]
Doench et al., “Supplementary Material: Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation.” Nature Biotechnology 32(12):1262-1267 (2014).
[cited by applicant]
Examination Search Report, May 11, 2018 in corresponding Canadian No. 2737180.
[cited by applicant]
Fenaux “Inhibitors of DNA methylation: beyond myelodysplastic syndromes.” Nature Reviews Clinical Oncology 2 (S21):S36-S44 (2005).
[cited by applicant]
Fischer et al., “Pulmonary Passage is a Major Obstacle for Intravenous Stem Cell Delivery: The Pulmonary First- Pass Effect” Stem Cells and Development 18(5):683-91 (2009).
[cited by applicant]
Flanagan et al., “Hydroxycarbamide alters erythroid gene expression in children with sickle cell anaemia.” British Journal of Haematology 157(2):240-248 (2012).
[cited by applicant]
GenBank Accession No. NM_022893.4. “
[cited by applicant]
GeneCard for BCL11A, retrieved from http://www.genecards.org/cgi-bin/carddisp.pl?gene=BCL11A on Jun. 22, 2012.
[cited by applicant]
Goffin et al., “DNA methyltransferase inhibitors—state of the art.” Annals of Oncology 13(11):1699-716 (2002).
[cited by applicant]
Goldberg et al., “Treatment of sickle cell anemia with hydroxyurea and erythropoietin.” New England Journal of Medicine 323(6):366-372 (1990).
[cited by applicant]
Hackam “Translating animal research into clinical benefit” BMJ 334:163-68 (2007).
[cited by applicant]
Hancarova et al. “A patient with de novo 0.45 Mb deletion of 2p16. 1: The role of BCL11A, PAPOLG, REL, and FLJ16341 in the 2p15-p16. 1 microdeletion syndrome.” American Journal of Medical Genetics Part A 161(4):865-870 …
[cited by applicant]
Harding et al., “Large animal models for stem cell therapy”, Stem Cell Research & Therapy 4(23):1-9 (2013).
[cited by applicant]
Hebbel et al., “The HDAC inhibitors trichostatin A and suberoylanilide hydroxamic acid exhibit multiple modalities of benefit for the vascular pathobiology of sickle transgenic mice.” Blood 115(12):2483-2490 (2010).
[cited by applicant]
Hendel et al., “Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells.” Nature Biotechnology 33(9):985-989 (2015).
[cited by applicant]
Higgs et al., “Genetic complexity in sickle cell disease.” PNAS 105(33):11595-11596 (2008).
[cited by applicant]
Ho et al., “In vitro induction of fetal hemoglobin in human erythroid progenitor cells.” Experimental Hematology 31 (7):586-591 (2003).
[cited by applicant]
Hsieh et al., “Allogeneic hematopoietic stem-cell transplantation for sickle cell disease.” New England Journal of Medicine 361(24):2309-2317 (2009).
[cited by applicant]
Jane et al., “Understanding fetal globin gene expression: a step towards effective HbF reactivation in haemoglobinopathies.” British Journal of Haematology 102(2):415-423 (1998).
[cited by applicant]
Kauf et al., “The cost of health care for children and adults with sickle cell disease.” American Journal of Hematology 84(6):323-327 (2009).
[cited by applicant]
Kirschner et al., “Genomic mapping of chromosomal region 2p15-p21 (D2S378-D2S391): integration of Genemap'98 within a framework of yeast and bacterial artificial chromosomes” Genomics 62(1):21-33 (1999).
[cited by applicant]
Boden et al. “Enhanced gene silencing of HIV-1 specific siRNA using microRNA designed hairpins.” Nucleic Acids Research 32(3): 1154-1158 (2004).
[cited by applicant]
Brendel et al. “Lineage-specific BCL11A knockdown circumvents toxicities and reverses sickle phenotype.” The Journal of Clinical Investigation 126(10): 3868-3878 (2016).
[cited by applicant]
Brendel et al. “Optimization of Bcl11a Knockdown by miRNA scaffold embedded shmas leading to enhanced induction of fetal hemoglobin in erythroid cells for the treatment of beta-hemoglobinopathies.” Blood 124(21): 2150-2…
[cited by applicant]
Calloni et al. “Scaffolds for artificial miRNA expression in animal cells.” Human Gene Therapy Methods 26(5): 162-174 (2015).
[cited by applicant]
Cante-Barrett et al. “Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters.” BMC Research Notes 9(1): 1-6 (2016).
[cited by applicant]
Cavazzana-Calvo et al. “Transfusion independence and HMGA2 activation after gene therapy of human β-thalassaemia.” Nature 467(7313): 318-322 (2010).
[cited by applicant]
Clever et al. “RNA secondary structure and binding sites for gag gene products in the 5′packaging signal of human immunodeficiency virus type 1.” Journal of Virology 69(4): 2101-2109 (1995).
[cited by applicant]
Cullen et al. “Regulatory pathways governing HIV-1 replication.” Cell 58(3): 423-426 (1989).
[cited by applicant]
Cullen. “Human immunodeficiency virus as a prototypic complex retrovirus.” Journal of Virology 65(3): 1053-1056 (1991).
[cited by applicant]
Database GenBank [Online] Mar. 3, 2015, Anonymous: “TPA:
[cited by applicant]
Extended European Search Report mailed on Jan. 12, 2021, for European Application No. 18775163.1, 11 pages.
[cited by applicant]
Ginn et al. “Gene therapy clinical trials worldwide to 2012—an update.” The Journal of Gene Medicine 15(2): 65-77 (2013).
[cited by applicant]
Huang et al. “Role of the hepatitis B virus posttranscriptional regulatory element in export of intronless transcripts.” Molecular and Cellular Biology 15(7): 3864-3869 (1995).
[cited by applicant]
Imren et al. “Permanent and panerythroid correction of murine β thalassemia by multiple lentiviral integration in hematopoietic stem cells.” Proceedings of the National Academy of Sciences 99(22): 14380-14385 (2002).
[cited by applicant]
International Search Report and Written Opinion mailed on Jun. 25, 2018, for International Application No. PCT/US2018/025165, 10 paqes.
[cited by applicant]
Kitowski, “A Lentiviral Vector Conferring Coregulated, Erythroid-Specific Expression of [gamma]-Globin and shRNA Sequences to BCL11A for the Treatment of Sickle Cell Disease,” Aug. 1, 2016, 105 pages.
[cited by applicant]
Kutner et al. “Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors.” Nature Protocols 4(4): 495-505 (2009).
[cited by applicant]
Kutner et al. “Simplified production and concentration of HIV-1-based lentiviral vectors using HYPERFlask vessels and anion exchange membrane chromatography.” BMC Biotechnology 9(1): 1-7 (2009).
[cited by applicant]
Landau et al. “Packaging system for rapid production of murine leukemia virus vectors with variable tropism.” Journal of Virology 66(8): 5110-5113 (1992).
[cited by applicant]
Liu et al. “HnRNP L binds a cis-acting RNA sequence element that enables intron-dependent gene expression.” Genes & Development 9(14): 1766-1780 (1995).
[cited by applicant]
Luc et al. “Bcl11a deficiency leads to hematopoietic stem cell defects with an aging-like phenotype.” Cell Reports 16 (12): 3181-3194 (2016).
[cited by applicant]
Mahajan et al. “Control of beta globin genes.” Journal of Cellular Biochemistry 102(4): 801-810 (2007).
[cited by applicant]
Malik et al. “Successful Correction of the Human Cooley's Anemia β-Thalassemia Major Phenotype Using a Lentiviral Vector Flanked by the Chicken Hypersensitive Site 4 Chromatin Insulator.” Annals of the New York Academy …
[cited by applicant]
May et al. “Therapeutic haemoglobin synthesis in β-thalassaemic mice expressing lentivirus-encoded human β-globin.” Nature 406(6791): 82-86 (2000).
[cited by applicant]
Naldini. “Gene therapy returns to centre stage.” Nature 526(7573): 351-360 (2015).
[cited by applicant]
Negre et al. “Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown, β-Globin Induction and Simultaneous Repression of β-Globin for the Potential Treatment of Sickle Cell Disease.…
[cited by applicant]
Pawliuk et al. “Correction of sickle cell disease in transgenic mouse models by gene therapy.” Science 294(5550): 2368-2371 (2001).
[cited by applicant]
Pawliuk et al. “Correction of sickle cell disease in transgenic mouse models by gene therapy.” Science 294(5550): 2368-2371 (2001) [Supplemental Material].
[cited by applicant]
Rasmussen et al. “The miR-144/451 locus is required for erythroid homeostasis.” Journal of Experimental Medicine 207(7): 1351-1358 (2010).
[cited by applicant]
Soneoka et al. “A transient three-plasmid expression system for the production of high titer retroviral vectors.” Nucleic Acids Research 23(4): 628-633 (1995).
[cited by applicant]
Wang et al. “A ‘suicide’ CRISPR-Cas9 system to promote gene deletion and restoration by electroporation in
[cited by applicant]
Wu et al. “Highly efficient therapeutic gene editing of human hematopoietic stem cells.” Nature Medicine 25(5): 776-783 (2019).
[cited by applicant]
Yu et al. “Bcl11a is essential for lymphoid development and negatively regulates p53.” Journal of Experimental Medicine 209(13): 2467-2483 (2012).
[cited by applicant]
Zennou et al. “HIV-1 genome nuclear import is mediated by a central DNA flap.” Cell 101(2): 173-185 (2000).
[cited by applicant]
Zufferey et al. “Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors.” Journal of Virology 73(4): 2886-2892 (1999).
[cited by applicant]
Bofill-De Ros et al. “Guidelines for the optimal design of miRNA-based shRNAs.” Methods 103: 157-166 (2016).
[cited by applicant]
Guda et al. “miRNA-embedded shRNAs for lineage-specific BCL11A knockdown and hemoglobin F induction.” Molecular Therapy 23(9): 1465-1474 (2015).
[cited by applicant]
Koshy et al., “2-deoxy 5-azacytidine and fetal hemoglobin induction in sickle cell anemia.” Blood 96(7):2379-2384 (2000).
[cited by applicant]
Labie “Le contrôle en trans de la production d'hémoglobine fœtale: une recherche qui dure depuis 20 ans.” Hématologie 14(2):165-166 (2008).
[cited by applicant]
Lettre et al., “DNA polymorphisms at the BCL11A, HBS1L-MYB, and beta-globin loci associate with fetal hemoglobin levels and pain crises in sickle cell disease.” PNAS 105(33):11869-11874 (2008).
[cited by applicant]
Liu et al., “Bcl11a is essential for normal lymphoid development.” Nature Immunology 4(6):525-532 (2003).
[cited by applicant]
Liu et al., “Functional studies of BCL11A: characterization of the conserved BCL11A-XL splice variant and its interaction with BCL6 in nuclear paraspeckles of germinal center B cells.” Molecular Cancer 5(18):1-6 (2006).
[cited by applicant]
Lulli et al., “MicroRNA-486-3p regulates γ-globin expression in human erythroid cells by directly modulating BCL11A.” PLoS One 8(4):e60436 (2013).
[cited by applicant]
Makala et al., “Fetal Hemoglobin Induction to Treat b-Hemoglobinopathies: From Bench to Bedside” J Hematol Transfus 2(2):1-2 (2014).
[cited by applicant]
Martin-Subero et al., “Recurrent involvement of the REL and BCL11Aloci in classical Hodgkin lymphoma.” Blood 99(4):1474-1477 (2002).
[cited by applicant]
Matsuda et al., “Transcription factors LRF and BCL11A independently repress expression of fetal hemoglobin” Science 351(6270):285-289 (2016).
[cited by applicant]
Menzel et al., “A QTL Influencing F cell production maps to a gene encoding a zinc-finger protein on chromosome 2p15.” Nature Genetics 39(10):1197-1199 (2007).
[cited by applicant]
Migliaccio et al., “Influence of recombinant hematopoietins and of fetal bovine serum on the globin synthetic pattern of human BFUe.” Blood 76(6):1150-1157 (1990).
[cited by applicant]
Moffat et al., “A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen.” Cell 124(6):1283-1298 (2006).
[cited by applicant]
Nemudryi et al., “TALEN and CRISPR/Cas Genome Editing Systems: Tools of Discovery” Acta Naturae, 6(3):19-40 (2014).
[cited by applicant]
Neven et al., “A Mendelian predisposition to B-cell lymphoma caused by IL-10R deficiency,” Blood 3713-3722 (2013).
[cited by applicant]
Orkin et al., “Recent advances in globin research using genome-wide association studies and gene editing.” Annals of the New York Academy of Sciences 1368 (1):5-10 (2016).
[cited by applicant]
Papayannopoulou et al., “Erythroid progenitors circulating in the blood of adult individuals produce fetal hemoglobin in culture.” Science 199(4335):1349-1350 (1978).
[cited by applicant]
Pauling et al., “Sickle cell anemia a molecular disease.” Science 110(2865):543-548 (1949).
[cited by applicant]
Pembrey et al., “Fetal haemoglobin production and the sickle gene in the oases of Eastern Saudi Arabia.” British journal of haematology 40(3):415-429 (1978).
[cited by applicant]
Perrine “Fetal globin induction—can it cure beta thalassemia?” American Society of Hematology Education Program Book pp. 38-44 (2005).
[cited by applicant]
Platt et al., “Mortality in sickle cell disease. Life expectancy and risk factors for early death.” New England Journal of Medicine 330(23):1639-1644 (1994).
[cited by applicant]
Purton et al., “All-trans retinoic acid enhances the long-term repopulating activity of cultured hematopoietic stem cells.” Blood 95(2):470-477 (2000).
[cited by applicant]
Renella et al. “Hematopoietic SIN lentiviral micro RNA-mediated silencing of BCL11A: pre-clinical evidence for a sickle cell disease gene-therapy trial.” Blood 120(21):Abstract 753 (2012).
[cited by applicant]
Ridley et al., “Erythropoietin: A Review” J Natl Med Assoc., 86(2):129-135 (1994).
[cited by applicant]
Rodriguez et al., “A bioavailability and pharmacokinetic study of oral and intravenous hydroxyurea.” Blood 9 (5):1533-1541 (1998).
[cited by applicant]
Roggenkamp et al. “Tuning CRISPR-Cas9 gene drives in
[cited by applicant]
Rosenblum et al., “Peripheral blood erythroid progenitors from patients with sickle cell anemia: HPLC separation of hemoglobins and the effect of a HbF switching factor.” Progress in Clinical and Biological Research 191…
[cited by applicant]
Saiki et al., “Human EVI9, a homologue of the mouse myeloid leukemia gene, is expressed in the hematopoletic progenitors and down-regulated during myeloid differentiation of HL60 cells.” Genomics 70(3):387-391 (2000).
[cited by applicant]
Sankaran et al., “Developmental and species-divergent globin switching are driven by BCL11A.” Nature 460 (7259):1093-1097 (2009).
[cited by applicant]
Sankaran et al., “Human fetal hemoglobin expression is regulated by the developmental stage-specific repressor BCL11A.” Science 322(5909):1839-1843 (2008).
[cited by applicant]
Sankaran et al., “Targeted therapeutic strategies for fetal hemoglobin induction.” American Society of Hematology Education Program Book 2011(1):459-465 (2011).
[cited by applicant]
Satterwhite et al., “The BCL11 gene family: involvement of BCL11A in lymphoid malignancies.” Blood, 98 (12):3413-3420 (2001).
[cited by applicant]
Schopman et al. “Optimization of shRNA inhibitors by variation of the terminal loop sequence.” Antiviral Research 86 (2):204-211 (2010).
[cited by applicant]
Sebastiani et al., “BCL11A enhancer haplotypes and fetal hemoglobin in sickle cell anemia.” Blood Cells, Molecules, and Diseases 54(3):224-230 (2015).
[cited by applicant]
Sedgewick et al., “BCL11A is a major HbF quantitative trait locus in three different populations with β-hemoglobinopathies.” Blood Cells, Molecules, and Diseases 41(3):255-258 (2008).
[cited by applicant]
Shen et al., “Modifcation of globin gene expression by RNA targeting strategies.” Experimental Hematology, 35 (8):1209-1218 (2007).
[cited by applicant]
Takeuchi et al., “Redesign of extensive protein-DNA interfaces of meganucleases using iterative cycles of in vitro compartmentalization.” PNAS 111(11):4061-4066 (2014).
[cited by applicant]
Taymans et al., “Radiation hybrid mapping of chromosomal region 2p15-p16: integration of expressed and polymorphic sequences maps at the Carney complex (CNC) and Doyne honeycomb retinal dystrophy (DHRD) loci.” Genomics …
[cited by applicant]
Terasawa et al., “Synthetic pre-miRNA-based shRNA as potent RNAi triggers.” Journal of Nucleic Acids (2011).
[cited by applicant]
Thein “Genetic modifiers of the beta-haemoglobinopathies.” British Journal of Hematology, 141(3):357-366 (2008).
[cited by applicant]
Thein et al., “Discovering the genetics underlying foetal haemoglobin production in adults.” British Journal of Haematology 145(4):455-467 (2009).
[cited by applicant]
Thompson “Structure, Function, and Molecular Defects of Factor IX.” Blood 67(3):565-72 (1986).
[cited by applicant]
Uda et al., “Genome-wide association study shows BCL11A associated with persistent fetal hemoglobin and amelioration of the phenotype of β-thalassemia.” PNAS 105(5):1620-1625 (2008).
[cited by applicant]
UCSC Genome Browser of Human Assembly, hg19. Feb. 2009. 1 Page.
[cited by applicant]
Wang et al. “Genetic screens in human cells using the CRISPR/Cas9 system.” Science 343(6166):80-84 (2013).
[cited by applicant]
Wang et al. “Supplementary Material: Genetic screens in human cells using the CRISPR/Cas9 system.” Science 343 (6166):80-84 (2013).
[cited by applicant]
Wang et al., “In Vivo Delivery Systems for Therapeutic Genome Editing” International Journal of Molecular Sciences 17(5):1-19 (2016).
[cited by applicant]
Wang et al., “Selection of hyperfunctional siRNAs with improved potency and specificity.” Nucleic Acids Research 37 (22):e152 (2009).
[cited by applicant]
White et al., “Factor VIII Gene and Hemophili A.” Blood 73(1):1-12 (1989).
[cited by applicant]
World Health Organization. “Sickle-cell anaemia. Report A59/9. Provisional agenda item 11.4.” 59th World Health Assembly. www.who.int/gb/ebwha/pdf_files/WHA59/A59_9-en.pdf (2006).
[cited by applicant]
Xu et al., “Correction of sickle cell disease in adult mice by interference with fetal hemoglobin silencing”, Science 334 (6058):993-996 (2011).
[cited by applicant]
Knowles et al. “Palmitate diet-induced loss of cardiac caveolin-3: a novel mechanism for lipid-induced contractile dysfunction.” PLoS One 8(4): e61369 pp. 1-11 (2013).
[cited by applicant]
Xu et al., “Reactivation of silenced human HbF in adult mice by inactivation of BCL11A.” Blood 116:Abstract 643 (2010).
[cited by applicant]
Xu et al., “Transcriptional silencing of beta-globin by BCL11A involvs long-range interactions and cooperation with SOX6.” Genes and Development 24(8):783-798 (2010).
[cited by applicant]
Yin et al. “BCL11A: a potential diagnostic biomarker and therapeutic target in human diseases.” Bioscience Reports 39(11): 1-13 (2019).
[cited by applicant]
Yin et al., “Bcl11a Causes p21 Cip1 Down-Regulation and Transplantable Leukemia in Nf1-Deficient Mice.” Blood 110 (11):2657-2657 (2007) [Abstract Only].
[cited by applicant]
Esrick et al. “Post-transcriptional genetic silencing of BCL11A to treat sickle cell disease.” New England Journal of Medicine 384(3): 205-215 (2021).
[cited by applicant]
Win et al. “A modular and extensible RNA-based gene-regulatory platform for engineering cellular function.” Proceedings of the National Academy of Sciences 104(36): 14283-14288 (2007).
[cited by applicant]
Johari et al. “Integrated cell and process engineering for improved transient production of a “difficult-to-express” fusion protein by CHO cells.” Biotechnology and Bioengineering 112(12): 2527-2542 (2015).
[cited by applicant]
Aouida et al. “Efficient fdCas9 synthetic endonuclease with improved specificity for precise genome engineering.” PloS one 10.7: e0133373 (2015).
[cited by applicant]
Chakraborty et al. “A CRISPR/Cas9-based system for reprogramming cell lineage specification.” Stem cell reports 3.6: 940-947 (2014).
[cited by applicant]
Chakraborty et al. “A CRISPR/Cas9-based system for reprogramming cell lineage specification.” Stem cell reports 3.6: supplementary information. (2014).
[cited by applicant]
Chang et al. “Genome editing with RNA-guided Cas9 nuclease in zebrafish embryos.” Cell research 23.4: 465-472 (2013).
[cited by applicant]
IDT. “Alt-R CRISPR-Cas9 System User Guide for Cationic Lipid Delivery of CRISPR Ribonucleoprotein into Mammalian Cells.” 2016.
[cited by applicant]
Miura et al. “An Efficient CRISPR protocol for generating Conditional and Knock-in mice using long single-stranded DNA donors.” bioRxiv: 141424 (2017).
[cited by applicant]
SCORE Sequence Alignment Results for Application Nos. PCT/US2019/030219 and U.S. Appl. No. 17/051,583, dated Jun. 6, 2024 (5 Pages).
[cited by applicant]