公开(公告)号：US20160304893A1

公开(公告)日：2016-10-20

申请号：US15103773

申请日：2014-12-12

Applicant: CELLECTIS

Inventor： Fayza DABOUSSI ,  Marine BEURDELEY ,  Alexandre JUILLERAT

IPC: C12N15/82 ,  C12N9/22

CPC classification number: C12N15/8213 ,  C12N9/22

Abstract: The present invention relates to a method of genome engineering in microalgae using the Cas9/CRISPR system. In particular, the present invention relates to methods of delivering RNA guides via cell penetrating peptides in microalgae, preferably in stable integrated Cas9 microalgae. The present invention also relates to kits and isolated cells comprising Cas9, split Cas9 or guide RNA and Cas9-fused cell-penetrating peptides. The present invention also relates to isolated cells obtained by the methods of the invention.

Abstract translation: 本发明涉及使用Cas9 / CRISPR系统的微藻中的基因组工程方法。 特别地，本发明涉及通过微藻中的细胞穿透肽递送RNA引导物的方法，优选在稳定的综合Cas9微藻中。 本发明还涉及包含Cas9，裂解Cas9或引导RNA和Cas9融合的细胞穿透肽的试剂盒和分离的细胞。 本发明还涉及通过本发明的方法获得的分离的细胞。

公开(公告)号：US20160273002A1

公开(公告)日：2016-09-22

申请号：US15031996

申请日：2014-10-24

Applicant: CELLECTIS

Inventor： Philippe DUCHATEAU ,  Alexandre JUILLERAT

IPC: C12N15/90 ,  C07K14/195 ,  C12N9/22

CPC classification number: C12N15/907 ,  A61K38/00 ,  A61K48/00 ,  A61K48/005 ,  C07K14/195 ,  C07K2319/80 ,  C12N9/22 ,  C12Y301/00 ,  C12Y301/21004

Abstract: The present invention is in the field of genetic editing tools and methods of genetic engineering. It relates to the engineering of rare-cutting endonucleases designed to contract highly repetitive motives in chromosomes, which are at the origin of certain genetic diseases, in particular the so-called “triplet repeat diseases”, such as the Huntington disease. The invention encompasses the method for contracting the repetitive motives, the rare-cutting endonucleases for use to contract repetitive motives in a gene subjected to repeat disorder, the polynucleotides and vectors encoding thereof as well as the resulting pharmaceutical compositions.

Abstract translation: 本发明在遗传编辑工具和基因工程方法领域。 它涉及设计用于在染色体中收缩高度重复动机的稀有内切核酸酶的工程，这些动机是某些遗传疾病的起源，特别是所谓的“三重重复疾病”，如亨廷顿疾病。 本发明包括用于收缩重复动机的方法，用于在重复病症的基因中重复动作的稀有内切核酸酶，编码它们的多核苷酸和载体以及所得药物组合物。

公开(公告)号：US20160184362A1

公开(公告)日：2016-06-30

申请号：US15045368

申请日：2016-02-17

Applicant: Cellectis

Inventor： Philippe DUCHATEAU ,  André CHOULIKA ,  Laurent POIROT

IPC: A61K35/17 ,  C12N9/22 ,  C12N15/90 ,  A61K47/48 ,  C12N5/0783

CPC classification number: C12N15/85 ,  A61K35/17 ,  C12N5/0636 ,  C12N5/0637 ,  C12N5/0638 ,  C12N9/22 ,  C12N15/1138 ,  C12N15/90 ,  C12N2310/20 ,  C12N2510/00 ,  C12Y301/00

Abstract: The present invention relates to methods of developing genetically engineered, preferably non-alloreactive T-cells for immunotherapy. This method involves the use of RNA-guided endonucleases, in particular Cas9/CRISPR system, to specifically target a selection of key genes in T-cells. The engineered T-cells are also intended to express chimeric antigen receptors (CAR) to redirect their immune activity towards malignant or infected cells. The invention opens the way to standard and affordable adoptive immunotherapy strategies using T-Cells for treating cancer and viral infections.

公开(公告)号：US20160102324A1

公开(公告)日：2016-04-14

申请号：US14892707

申请日：2014-05-28

Applicant: CELLECTIS

Inventor： Philippe DUCHATEAU ,  Claudia BERTONATI

IPC: C12N15/90 ,  C12N9/22

CPC classification number: C12N15/907 ,  C12N9/22 ,  C12N15/1082 ,  C12N15/63 ,  C12N15/902 ,  C12Y301/00 ,  C12Y301/26004

Abstract: The present invention is in the field of CRISPR-Cas system for genome targeting. The present invention relates to new engineered Cas9 scaffolds and uses thereof. More particularly, the present invention relates to methods for genome targeting, cell engineering and therapeutic application. The present invention also relates to vectors, compositions and kits in which the new Cas9 scaffolds of the present invention are used.

Abstract translation: 本发明涉及用于基因组靶向的CRISPR-Cas系统。 本发明涉及新的工程化Cas9支架及其用途。 更具体地，本发明涉及用于基因组靶向，细胞工程和治疗应用的方法。 本发明还涉及其中使用本发明的新的Cas9支架的载体，组合物和试剂盒。

公开(公告)号：US20150315557A1

公开(公告)日：2015-11-05

申请号：US14744668

申请日：2015-06-19

Applicant: CELLECTIS

Inventor： Andre Choulika ,  Frederic Cedrone ,  Julianne Smith

IPC: C12N9/22

CPC classification number: C12N9/22 ,  A61K38/00 ,  C12N2710/16622 ,  C12N2730/10122 ,  Y02A50/467

Abstract: An I-CreI variant, wherein at least one of the two I-Cre1 monomers has at least two substitutions, one in each of the two functional subdomains of the LAGLIDADG core domain situated from positions 26 to 40 and 44 to 77 of I-CreI, said variant being able to cleave a DNA target sequence from the genome of a non-integrating virus, in particular herpes simplex virus (HSV) or Hepatitis B virus (HBV) for use in genome engineering and for in vivo and ex vivo (gene cell therapy) genome therapy as well as the treatment of a virus infection.

Abstract translation: I-CreI变体，其中两个I-Cre1单体中的至少一个具有至少两个取代，位于位于I-CreI的位置26至40和44至77的LAGLIDADG核心结构域的两个功能亚结构域的每一个中的一个 所述变体能够从非整合型病毒，特别是单纯疱疹病毒（HSV）或乙型肝炎病毒（HBV）的基因组中切割DNA靶序列用于基因组工程和体内和离体（基因 细胞治疗）基因组治疗以及病毒感染的治疗。

公开(公告)号：US20150272076A1

公开(公告)日：2015-10-01

申请号：US14439420

申请日：2013-10-31

Applicant: CELLECTIS SA ,  MEDICAGO INC.

Inventor： Luc Mathis ,  Daniel F. Voytas ,  Jin Li ,  Feng Zhang ,  Thomas Stoddard ,  Marc-Andre D'Aoust

IPC: A01H5/12 ,  A01H1/06 ,  C12P21/00

CPC classification number: A01H5/12 ,  A01H1/06 ,  C12N9/22 ,  C12N15/01 ,  C12N15/8218 ,  C12N15/8257 ,  C12N15/8258 ,  C12P21/00 ,  C12P21/005

Abstract: Materials and methods are provided for making plants (e.g., Nicotiana varieties) that are suitable for producing therapeutic polypeptides suitable for administration to humans and animals, particularly by making TAL effector endonuclease-induced mutations in genes encoding xylosyltransferases and fucosyltransferases.

Abstract translation: 提供材料和方法用于制造适合于产生适于施用于人和动物的治疗性多肽的植物（例如烟草属），特别是通过在编码木糖基转移酶和岩藻糖基转移酶的基因中产生TAL效应子内切核酸酶诱导的突变。

公开(公告)号：US20150218230A1

公开(公告)日：2015-08-06

申请号：US14424757

申请日：2013-08-30

Applicant: CELLECTIS, S.A.

Inventor： Alexandre Juillerat ,  Philippe Duchateau

IPC: C07K14/195 ,  C07H21/00

CPC classification number: C07K14/195 ,  C07H21/00 ,  C07K2319/80 ,  C12N15/10 ,  C12N15/102 ,  C12N15/63 ,  C12N15/8216 ,  C12N2501/60

Abstract: Methods for improving or modulating targeting specificity of TALE proteins by introducing alternative RVDs into their modular nucleic acid binding domains. Polynucleotides encoding TALE proteins having alternative targeting specificity towards a nucleic acid target sequence. TALE proteins having alternative targeting specificity towards a nucleic acid target sequence and methods of making and using them.

Abstract translation: 通过将替代RVD引入其模块化核酸结合结构域来改善或调节TALE蛋白的靶向特异性的方法。 编码具有针对核酸靶序列的替代靶向特异性的TALE蛋白的多核苷酸。 具有针对核酸靶序列的替代靶向特异性的TALE蛋白质及其制备和使用它们的方法。

公开(公告)号：US20150203871A1

公开(公告)日：2015-07-23

申请号：US14405283

申请日：2013-06-05

Applicant: Cellectis

Inventor： Alexandre Juillerat ,  Philippe Duchateau

IPC: C12N15/90

CPC classification number: C12N15/907 ,  C07K2319/80 ,  C12N9/22 ,  C12N15/10

Abstract: The present invention relates to Transcription Activator-Like Effector (TALE) derived proteins that allow to efficiently target and/or process double stranded nucleic acid sequences. The proteins of the invention are typically chimeric protein monomers composed of a core scaffold comprising Repeat Variable Dipeptide regions (RVDs) having binding specificity to a DNA target sequence, to which is fused a catalytic domain to its N-terminal. This later catalytic domain, which can be a monomer of a nuclease, is placed at this position to possibly interact with another catalytic domain fused to another TAL monomer, such that, when said monomers are binding to their respective target DNA sequences, both catalytic domains form a catalytic entity likely to process DNA in the proximity of these target sequences. This new TAL architecture makes it possible to target only one DNA strand, which is not the case, for instance, with classical TALEN architectures. The present invention also relates to vectors encoding such proteins and compositions or kits in which Transcription Activator-Like Effector (TALE) proteins of the present invention are used.

Abstract translation: 本发明涉及允许有效靶向和/或加工双链核酸序列的转录激活子样效应子（TALE）衍生蛋白。 本发明的蛋白质通常是由包含对DNA靶序列具有结合特异性的重复可变二肽区域（RVD）的核心支架构成的嵌合蛋白质单体，将催化结构域融合到其N-末端。 该后续的催化结构域（其可以是核酸酶的单体）置于该位置以可能与另一个与另一个TAL单体融合的另一催化结构域相互作用，使得当所述单体与其各自的靶DNA序列结合时，两个催化结构域 形成可能在这些靶序列附近处理DNA的催化实体。 这种新的TAL架构使得可以仅靶向一条DNA链，而不是例如经典的TALEN架构。 本发明还涉及编码这样的蛋白质的载体，其中使用本发明的转录激活剂样效应子（TALE）蛋白质的组合物或试剂盒。

公开(公告)号：US20140178942A1

公开(公告)日：2014-06-26

申请号：US14064775

申请日：2013-10-28

Applicant: CELLECTIS

Inventor： ANDRE CHOULIKA ,  ROMAN GALETTO

IPC: C12P19/34

CPC classification number: C12P19/34 ,  A61K38/00 ,  C12N9/22 ,  C12N2740/16022

Abstract: Meganuclease variants which cleave at least one target in the provirus of a retrovirus and in particular which cleave the genomic insertion of the provirus. The present invention particular relates to meganuclease variants which cleave the provirus of the Human Immunodeficiency Virus genome following genomic insertion. Vector encoding such variants, as well as to a cell or multi-cellular organism modified by such a vector and use of said meganuclease variants and derived products for genome engineering and for in vivo and ex vivo (gene cell therapy) genome therapy.

Abstract translation: 在逆转录病毒的前病毒中切割至少一个靶，特别是切割前病毒的基因组插入的大范围核酸酶变体。 本发明特别涉及在基因组插入后切割人免疫缺陷病毒基因组的原病毒的大范围核酸酶变体。 编码这样的变体的载体，以及通过这种载体修饰的细胞或多细胞生物体，以及使用所述大范围核酸酶变体和衍生产物用于基因组工程和用于体内和离体（基因细胞治疗）基因组治疗。

公开(公告)号：US20130209437A1

公开(公告)日：2013-08-15

申请号：US13744068

申请日：2013-01-17

Applicant: Cellectis

Inventor： Frederic Paques

IPC: C12N9/22 ,  C12N15/10 ,  C12N15/90 ,  A01N63/00

CPC classification number: C12N9/22 ,  A01N63/00 ,  C12N15/1086 ,  C12N15/902

Abstract: A method for engineering I-CreI homing endonuclease variants able to cleave mutant I-CreI sites having variation in positions ±8 to ±10. A I-CreI homing endonuclease variant obtainable by said method, a vector encoding said variant, a cell, an animal or a plant modified by said vector. Use of said I-CreI endonuclease variant and derived products for genetic engineering, genome therapy and antiviral therapy.

Abstract translation: 用于工程化I-CreI归巢内切核酸酶变体的方法，其能够切割具有±8至±10位置变异的突变I-CreI位点。 通过所述方法可获得的I-CreI归巢内切核酸酶变体，编码所述变体的载体，细胞，动物或由所述载体修饰的植物。 使用所述I-CreI内切核酸酶变体和衍生产物用于基因工程，基因组治疗和抗病毒治疗。