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1. 发明申请

US20200318110A1 A COMPOSITION AND METHOD OF USING MIR-302 PRECURSORS AS ANTI-CANCER DRUGS FOR TREATING HUMAN LUNG CANCER 审中-公开
公开(公告)号：US20200318110A1
公开(公告)日：2020-10-08
申请号：US16305030
申请日：2017-02-24
申请人： Shi-Lung LIN , David TS WU
发明人： Shi-Lung LIN , David TS WU , Hsuan-Hsuan LU
IPC分类号： C12N15/113 , A61K31/7105 , A61P35/04
摘要： This invention generally relates to a composition and method of using man-made small RNAs, such as small interfering RNAs (siRNA), microRNAs (miRNA) and their hairpin-like precursors (pre-miRNA), as tumor suppressing anti-cancer drugs for treating human tumors and cancers, in particular, but not limited, for treating skin (melanoma), blood (leukemia), prostate, breast, liver and lung cancers as well as various neoplastic tumors, such as brain tumors and teratocarcinomas that contain a variety of tumorous and cancerous cells derived from all three germ layers of tissues, including ectoderm, mesoderm and endoderm. More specifically, the present invention relates to the use of miR-302-like siRNA (siR-302) and/or miR-302 precursors (pre-miR-302) for developing novel medicines and therapies against a variety of human cancers, in particular lung cancers.

2. 发明申请

US20170175118A1 COMPOSITION AND METHOD OF USING MIR-302 PRECURSORS AS ANTI-CANCER DRUGS FOR TREATING HUMAN LUNG CANCER 审中-公开
公开(公告)号：US20170175118A1
公开(公告)日：2017-06-22
申请号：US15442557
申请日：2017-02-24
申请人： Shi-Lung Lin , Donald C. Chang , David TS Wu , Hsuan-Hsuan Lu
发明人： Shi-Lung Lin , Donald C. Chang , David TS Wu , Hsuan-Hsuan Lu
IPC分类号： C12N15/113
CPC分类号： C12N15/113 , C12N15/111 , C12N15/1135 , C12N2310/14 , C12N2310/141 , C12N2310/531 , C12N2320/30 , C12N2320/32 , C12N2330/50 , C12N2330/51
摘要： This invention generally relates to a composition and method of using mam-made small RNAs, such as small interfering RNAs (siRNA), microRNAs (miRNA) and their hairpin-like precursors (pre-miRNA), as tumor suppressing anti-cancer drugs for treating human tumors and cancers, in particular, but not limited, for treating skin (melanoma), blood (leukemia), prostate, breast, liver and lung cancers as well as various neoplastic tumors, such as brain tumors and teratocarcinomas that contain a variety of tumorous and cancerous cells derived from all three germ layers of tissues, including ectoderm, mesoderm and endoderm. More specifically, the present invention relates to the use of miR-302-like siRNA (siR-302) and/or miR-302 precursors (pre-miR-302) for developing novel medicines and therapies against a variety of human cancers, in particular lung cancers.

3. 发明申请

US20200165607A1 COMPOSITION AND METHOD OF USING MIR-302 PRECURSORS AS ANTI-CANCER DRUGS FOR TREATING HUMAN LUNG CANCER 审中-公开
公开(公告)号：US20200165607A1
公开(公告)日：2020-05-28
申请号：US16532353
申请日：2019-08-05
申请人： Shi-Lung Lin , Donald C. Chang , David TS Wu , Hsuan-Hsuan Lu
发明人： Shi-Lung Lin , Donald C. Chang , David TS Wu , Hsuan-Hsuan Lu
IPC分类号： C12N15/113 , C12N15/11
摘要： This invention generally relates to a composition and method of using mam-made small RNAs, such as small interfering RNAs (siRNA), microRNAs (miRNA) and their hairpin-like precursors (pre-miRNA), as tumor suppressing anti-cancer drugs for treating human tumors and cancers, in particular, but not limited, for treating skin (melanoma), blood (leukemia), prostate, breast, liver and lung cancers as well as various neoplastic tumors, such as brain tumors and teratocarcinomas that contain a variety of tumorous and cancerous cells derived from all three germ layers of tissues, including ectoderm, mesoderm and endoderm. More specifically, the present invention relates to the use of miR-302-like siRNA (siR-302) and/or miR-302 precursors (pre-miR-302) for developing novel medicines and therapies against a variety of human cancers, in particular lung cancers.

4. 发明申请

US20160264974A1 USE OF MICRORNA PRECURSORS AS DRUGS FOR INDUCING CD34-POSITIVE ADULT STEM CELL EXPANSION 有权
标题翻译：使用MICRORNA前列腺素作为诱导CD34阳性成年干细胞扩增的药物
公开(公告)号：US20160264974A1
公开(公告)日：2016-09-15
申请号：US15167226
申请日：2016-05-27
申请人： Shi-Lung LIN , Donald CHANG , David TS WU
发明人： Shi-Lung LIN , Donald CHANG , David TS WU
IPC分类号： C12N15/113 , C12P19/34 , C12N15/70
CPC分类号： C12N15/1135 , C12N15/111 , C12N15/113 , C12N15/70 , C12N2310/14 , C12N2310/141 , C12N2320/30 , C12N2330/50 , C12N2330/51 , C12P19/34
摘要： This invention generally relates to a composition and its production method useful for developing drugs/vaccines and/or therapies against a variety of degenerative diseases in humans. Particularly, the present invention teaches the essential steps of production and purification processes necessary for producing small hairpin-like RNA (shRNA) compositions, such as microRNA precursors (pre-miRNA) and short interfering RNAs (siRNA), which are useful for treating human ageing related diseases, such as, but not limited, Alzheimer's diseases, Parkinson's diseases, osteoporosis, diabetes, and cancers. The novelty of the present invention is to create an artificially enhanced adaptation environment for prokaryotic cells to adopt eukaryotic pol-2 and/or pol-2-like promoters for transcribing desired ncRNAs and/or their precursors without going through error-prone prokaryotic promoters, so as to improve the productive efficiency and reading fidelity of the shRNA transcription in the prokaryotic cells. The resulting shRNAs, preferably pre-miRNAs and siRNAs, are useful for developing therapeutic drugs against human degenerative diseases, particularly through a mechanism to induce CD34-positive stem cell expansion and/or regeneration.
摘要翻译：本发明一般涉及用于开发针对人类各种退行性疾病的药物/疫苗和/或疗法的组合物及其制备方法。特别地，本发明教导了用于生产小发夹样RNA（shRNA）组合物，例如可用于治疗人类的微小RNA前体（pre-miRNA）和短干扰RNA（siRNA））所需的生产和纯化过程的基本步骤衰老相关疾病，例如但不限于阿尔茨海默病，帕金森病，骨质疏松症，糖尿病和癌症。本发明的新颖之处在于为原核细胞产生人造增强的适应环境，以采用真核的pol-2和/或pol-2样启动子来转录所需的ncRNA和/或其前体而不经历易错的原核启动子，从而提高原核细胞中shRNA转录的生产效率和阅读保真度。产生的shRNA，优选pre-miRNAs和siRNAs可用于开发针对人退行性疾病的治疗药物，特别是通过诱导CD34阳性干细胞扩增和/或再生的机制。

5. 发明申请

US20090203141A1 Generation of tumor-free embryonic stem-like pluripotent cells using inducible recombinant RNA agents 审中-公开
标题翻译：使用可诱导的重组RNA制剂产生无肿瘤的胚胎干样多能细胞
公开(公告)号：US20090203141A1
公开(公告)日：2009-08-13
申请号：US12318806
申请日：2009-01-08
申请人： Shi-Lung Lin , David TS Wu
发明人： Shi-Lung Lin , David TS Wu
IPC分类号： C12N15/86 , C12N15/74
CPC分类号： C12N15/63 , C12N2840/102 , C12N2840/44 , C12N2840/445
摘要： The present invention generally relates to a method for developing, generating and selecting tumor-free embryonic stem (ES)-like pluripotent cells using electroporation delivery of an inducible tumor suppressor mir-302 agent into mammalian cells. More particularly, the present invention relates to a method and composition for generating a Tet-On/Off recombinant transgene capable of expressing a manually re-designed mir-302 microRNA (miRNA)/shRNA agent under the control of doxycyclin (Dox) in human somatic/cancer cells and thus inducing certain specific gene silencing effects on the differentiation-associated genes and oncogenes of the cells, resulting in reprogramming the cells into an ES-like pluripotent state.
摘要翻译：本发明一般涉及使用诱导型抑制剂mir-302试剂向哺乳动物细胞中电穿孔递送来开发，产生和选择无肿瘤胚胎干（ES）样多能细胞的方法。更具体地说，本发明涉及一种能够在人的多西环素（Dox）控制下表达手工重新设计的mir-302微小RNA（miRNA）/ shRNA试剂的Tet-On / Off重组转基因的方法和组合物。体细胞/癌细胞，从而对细胞的分化相关基因和癌基因诱导某些特异性基因沉默效应，导致将细胞重新编程成ES样多能状态。

6. 发明申请

US20090170204A1 Novel cosmetic designs and products using intronic RNA 有权
标题翻译：新型化妆品设计和使用内含子RNA的产品
公开(公告)号：US20090170204A1
公开(公告)日：2009-07-02
申请号：US12003662
申请日：2007-12-28
申请人： Shi-Lung Lin , David Ts Wu
发明人： Shi-Lung Lin , David Ts Wu
IPC分类号： C07H21/02 , C12N15/64
CPC分类号： C12N15/111 , C12N15/113 , C12N2310/111 , C12N2310/141 , C12N2320/30 , C12N2320/50
摘要： The present invention relates to a method and composition for generating a non-naturally occurring intron and its components capable of being processed into small hairpin RNA (shRNA) and/or microRNA (miRNA) molecules by skin cells and thus inducing specific gene silencing effects on skin pigment-related genes and/or aging-causing genes in the cells. The gene silencing effects so obtained are not only useful for lightening and whitening skin colors but also useful for suppressing unwanted aging gene activities in skins.
摘要翻译：本发明涉及一种用于产生非天然存在的内含子及其组分的方法和组合物，其能够被皮肤细胞加工成小发夹RNA（shRNA）和/或微小RNA（miRNA）分子，从而诱导特定的基因沉默效应皮肤色素相关基因和/或衰老基因。如此获得的基因沉默效应不仅可用于减轻和增白皮肤颜色，还可用于抑制皮肤中不需要的老化基因活性。

7. 发明授权

US09453219B2 Cosmetic designs and products using intronic RNA 有权
标题翻译：使用内含子RNA的化妆品设计和产品
公开(公告)号：US09453219B2
公开(公告)日：2016-09-27
申请号：US12003662
申请日：2007-12-28
申请人： Shi-Lung Lin , David TS Wu
发明人： Shi-Lung Lin , David TS Wu
IPC分类号： C12N15/113 , C12N15/11
CPC分类号： C12N15/111 , C12N15/113 , C12N2310/111 , C12N2310/141 , C12N2320/30 , C12N2320/50
摘要： The present invention relates to a method and composition for generating a non-naturally occurring intron and its components capable of being processed into small hairpin RNA (shRNA) and/or microRNA (miRNA) molecules by skin cells and thus inducing specific gene silencing effects on skin pigment-related genes and/or aging-causing genes in the cells. The gene silencing effects so obtained are not only useful for lightening and whitening skin colors but also useful for suppressing unwanted aging gene activities in skins.
摘要翻译：本发明涉及一种用于产生非天然存在的内含子及其组分的方法和组合物，其能够被皮肤细胞加工成小发夹RNA（shRNA）和/或微小RNA（miRNA）分子，从而诱导特定的基因沉默效应皮肤色素相关基因和/或衰老基因。如此获得的基因沉默效应不仅可用于减轻和增白皮肤颜色，还可用于抑制皮肤中不需要的老化基因活性。

8. 发明申请

US20220396798A1 NOVEL MRNA COMPOSITION AND PRODUCTION METHOD FOR USE IN ANTI-VIRAL AND ANTI-CANCER VACCINES 有权
公开(公告)号：US20220396798A1
公开(公告)日：2022-12-15
申请号：US17489357
申请日：2021-09-29
申请人： Shi-Lung LIN , Samantha CHANG-LIN , Jack SK CHEN , David TS WU , Chia-Ning SHEN , Mei-Jung WANG
发明人： Shi-Lung LIN , Samantha CHANG-LIN , Jack SK CHEN , David TS WU , Chia-Ning SHEN , Mei-Jung WANG
IPC分类号： C12N15/113 , C12N15/85
摘要： This invention relates to a novel mRNA composition and its production method useful for developing and manufacturing RNA-based anti-viral and/or anti-cancer vaccines and medicines. This invention includes two types of mRNA constructs, namely “5′-hairpin messenger RNA (5hmRNA)” and “messenger-hairpin-messenger RNA (mhmRNA)”, respectively. Both of 5hmRNA and mhmRNA contain at least a hairpin-like stem-loop RNA structure. The 5hmRNA contains at least a stem-loop RNA structure in the 5′-UTR of a protein/peptide-coding mRNA, while the mhmRNA contains a middle stem-loop structure flanked with two protein/peptide-coding mRNA sequences on both sides. In mhmRNA, the first 5′-mRNA preferably encodes an RNA replicase, for amplifying the second 3′-mRNA in transfected cells. After transfection into target cells, 5hmRNA and mhmRNA can be further translated into at least a desired protein/peptide. To produce highly structured 5hmRNA and mhmRNA, a novel PCR-IVT methodology has been developed and used with a specially designed RNA polymerase-helicase mixture reaction

9. 发明申请

US20170342418A1 USE OF MICRORNA PRECURSORS AS DRUGS FOR INDUCING CD34-POSITIVE ADULT STEM CELL EXPANSION 审中-公开
公开(公告)号：US20170342418A1
公开(公告)日：2017-11-30
申请号：US15661346
申请日：2017-07-27
申请人： Shi-Lung LIN , Donald CHANG , David TS WU
发明人： Shi-Lung LIN , Donald CHANG , David TS WU
IPC分类号： C12N15/113 , C12P19/34 , C12N15/11 , C12N15/70
CPC分类号： C12N15/1135 , C12N15/111 , C12N15/113 , C12N15/70 , C12N2310/14 , C12N2310/141 , C12N2320/30 , C12N2330/50 , C12N2330/51 , C12P19/34
摘要： This invention generally relates to a composition and its production method useful for developing drugs/vaccines and/or therapies against a variety of degenerative diseases in humans. Particularly, the present invention teaches the essential steps of production and purification processes necessary for producing small hairpin-like RNA (shRNA) compositions, such as microRNA precursors (pre-miRNA) and short interfering RNAs (siRNA), which are useful for treating human ageing related diseases, such as, but not limited, Alzheimer's diseases, Parkinson's diseases, osteoporosis, diabetes, and cancers. The novelty of the present invention is to create an artificially enhanced adaptation environment for prokaryotic cells to adopt eukaryotic pol-2 and/or pol-2-like promoters for transcribing desired ncRNAs and/or their precursors without going through error-prone prokaryotic promoters, so as to improve the productive efficiency and reading fidelity of the shRNA transcription in the prokaryotic cells. The resulting shRNAs, preferably pre-miRNAs and siRNAs, are useful for developing therapeutic drugs against human degenerative diseases, particularly through a mechanism to induce CD34-positive stem cell expansion and/or regeneration.

10. 发明授权

US09567591B2 Generation of human embryonic stem-like cells using intronic RNA 有权
标题翻译：使用内含子RNA产生人胚胎干细胞样细胞
公开(公告)号：US09567591B2
公开(公告)日：2017-02-14
申请号：US12149725
申请日：2008-05-07
申请人： Shi-Lung Lin , Shao-Yao Ying , David Ts Wu
发明人： Shi-Lung Lin , Shao-Yao Ying , David Ts Wu
IPC分类号： C12N15/11 , C07H21/02 , C07H21/04 , C12Q1/68 , C12N5/02 , C12N15/64 , C12N15/63
CPC分类号： C12N15/63
摘要： This invention generally relates to a method for developing, generating and selecting human embryonic stem (hES)-like pluripotent cells using transgenic expression of intronic microRNA-like RNA agents. More particularly, the present invention relates to a method and composition for generating a non-naturally occurring intron and its intronic components capable of being processed into mir-302-like RNA molecules in mammalian cells and thus inducing certain specific gene silencing effects on differentiation-related and fate-determinant genes of the cells, resulting in reprogramming the cells into a pluripotent embryonic stem (ES)-cell-like state. The ES-like cells so obtained are strongly express hES cell markers, such as Oct3/4, SSEA-3 and SSEA-4, and can be guided into various tissue cell types by treating certain hormones and/or growth factors under a feeder-free cell culture condition in vitro, which may be used for transplantation and gene therapies. Therefore, the present invention offers a simple, effective and safe gene manipulation approach for not only reprogramming somatic cells into ES-like pluripotent cells but also facilitating the maintenance of pluripotent and renewal properties of ES cells under a feeder-free cell culture condition, preventing the tedious retroviral insertion of four large transcription factor genes into one single cell as used in the previous iPS methods.
摘要翻译：本发明一般涉及使用内含子微RNA样RNA试剂的转基因表达来开发，产生和选择人胚胎干（hES）样多能细胞的方法。更具体地说，本发明涉及一种用于产生非天然存在的内含子及其内含子组分的方法和组合物，其能够在哺乳动物细胞中加工成mir-302样RNA分子，从而诱导对分化 - 相关和命运决定基因，导致将细胞重编程为多能胚胎干（ES）细胞样状态。如此获得的ES样细胞强烈表达hES细胞标志物，例如Oct3 / 4，SSEA-3和SSEA-4，并且可以通过在饲养层上处理某些激素和/或生长因子而被引导到各种组织细胞类型中，体外免疫细胞培养条件，可用于移植和基因治疗。因此，本发明提供了一种简单，有效和安全的基因操作方法，用于不仅将体细胞重编程为ES样多能细胞，而且有助于在无饲养细胞培养条件下维持ES细胞的多能性和更新性质，预防将以前的iPS方法中使用的四个大转录因子基因的繁琐的逆转录病毒插入一个单细胞中。

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