公开(公告)号：US20230009208A1

公开(公告)日：2023-01-12

申请号：US17848523

申请日：2022-06-24

申请人： The Brigham and Women’s Hospital, Inc. ,  President and Fellows of Harvard College ,  Vilnius University

发明人： Joseph Italiano ,  Linas Mazutis ,  Jonathan N. Thon ,  David A. Weitz

IPC分类号： B01L3/00 ,  C12M3/06 ,  C12M1/12 ,  C12M1/34 ,  G01N33/86 ,  C12M1/00

摘要： A system and method are provided for harvesting target biological substances. The system includes a substrate and a first and second channel formed in the substrate. The channels longitudinally extending substantially parallel to each other. A series of gaps extend from the first channel to the second channel to create a fluid communication path passing between a series of columns with the columns being longitudinally separated by a predetermined separation distance. The system also includes a first source configured to selectively introduce into the first channel a first biological composition at a first channel flow rate and a second source configured to selectively introduce into the second channel a second biological composition at a second channel flow rate. The sources are configured to create a differential between the first and second channel flow rates to generate physiological shear rates along the second channel that are bounded within a predetermined range.

公开(公告)号：US11326196B2

公开(公告)日：2022-05-10

申请号：US16069404

申请日：2017-01-10

申请人： VILNIUS UNIVERSITY ,  ETH ZURICH

发明人： Linas Mazutis ,  Greta Stonyte ,  Vaidotas Kiseliovas ,  Rapolas Zilionis ,  Arvydas Janulaitis ,  Robertas Galinis ,  Sabine Studer ,  Donald Hilvert

IPC分类号： C12P19/34 ,  B01J19/00 ,  A61K9/50 ,  B01L3/00

摘要： Disclosed is a system and method for production of DNA particles and use thereof. The DNA particles can be produced by amplification of nucleic acid molecule(s). Alternatively, DNA particles can be prepared by condensing multiple DNA molecules. The DNA condensation into a particle is mainly triggered by pyrophosphate and positively charged cations (e.g. magnesium). DNA particles can be applied for numerous biological applications but not limited to directed evolution, proteomics, drug delivery and imaging. DNA particles can be used to synthesize proteins using in vitro transcription/translation reaction.

公开(公告)号：US20210379555A1

公开(公告)日：2021-12-09

申请号：US17323902

申请日：2021-05-18

申请人： President and Fellows of Harvard College ,  Vilnius University

发明人： David A. Weitz ,  Allon Moshe Klein ,  Ilke Akartuna ,  Linas Mazutis ,  Marc W. Kirschner

IPC分类号： B01J19/00 ,  B01L3/00 ,  B01F13/00 ,  C12Q1/6806

摘要： The present invention generally relates to microfluidics and labeled nucleic acids. For example, certain aspects are generally directed to systems and methods for labeling nucleic acids within microfluidic droplets. In one set of embodiments, the nucleic acids may include “barcodes” or unique sequences that can be used to distinguish nucleic acids in a droplet from those in another droplet, for instance, even after the nucleic acids are pooled together. In some cases, the unique sequences may be incorporated into individual droplets using particles and attached to nucleic acids contained within the droplets (for example, released from lysed cells). In some cases, the barcodes may be used to distinguish tens, hundreds, or even thousands of nucleic acids, e.g., arising from different cells or other sources.

公开(公告)号：US10385336B2

公开(公告)日：2019-08-20

申请号：US15448818

申请日：2017-03-03

申请人： VILNIUS UNIVERSITY

发明人： Virginijus Siksnys ,  Migle Kazlauskiene ,  Gintautas Tamulaitis

IPC分类号： C12N15/11 ,  C12N15/113 ,  C12N9/22 ,  C07K14/315

摘要： A Type III-A CRISPR-Cas (StCsm) complex of Streptococcus thermophilus comprising crRNA, Csm4, and Csm3 and use for cleavage of RNA bearing a nucleotide sequence complementary to the crRNA, in vitro or in vivo. Methods for site-specific cleavage/shredding of a target RNA molecule using an RNA-guided RNA endonuclease comprising a minimal complex of crRNA, Csm4, and Csm3, and methods of RNA knock-down and RNA knock-out are disclosed.

公开(公告)号：US20180304222A1

公开(公告)日：2018-10-25

申请号：US15991600

申请日：2018-05-29

申请人： President and Fellows of Harvard College ,  Vilnius University

发明人： David A. Weitz ,  Allon Moshe Klein ,  Ilke Akartuna ,  Linas Mazutis ,  Marc W. Kirschner

IPC分类号： B01J19/00 ,  B01F13/00

CPC分类号： B01J19/0046 ,  B01F13/0062 ,  B01J2219/00585 ,  B01J2219/00722 ,  B01L3/502761 ,  B01L3/502776 ,  B01L3/502784 ,  B01L7/52 ,  B01L2200/0652 ,  B01L2300/021 ,  B01L2300/0663 ,  B01L2300/0858 ,  B01L2300/0867 ,  B01L2300/0883 ,  C12Q1/6806 ,  C12Q2563/149 ,  C12Q2563/159 ,  C12Q2565/514

摘要： The present invention generally relates to microfluidics and labeled nucleic acids. For example, certain aspects are generally directed to systems and methods for labeling nucleic acids within microfluidic droplets. In one set of embodiments, the nucleic acids may include “barcodes” or unique sequences that can be used to distinguish nucleic acids in a droplet from those in another droplet, for instance, even after the nucleic acids are pooled together. In some cases, the unique sequences may be incorporated into individual droplets using particles and attached to nucleic acids contained within the droplets (for example, released from lysed cells). In some cases, the barcodes may be used to distinguish tens, hundreds, or even thousands of nucleic acids, e.g., arising from different cells or other sources.

公开(公告)号：US20180251814A1

公开(公告)日：2018-09-06

申请号：US15559689

申请日：2016-03-18

申请人： VILNIUS UNIVERSITY

发明人： Saulius KLIMASAUSKAS ,  Giedrius VILKAITIS ,  Milda MICKUTE

IPC分类号： C12Q1/6806 ,  C12Q1/6853 ,  C12Q1/6876

CPC分类号： C12Q1/6806 ,  C12Q1/6853 ,  C12Q1/6876 ,  C12Q2521/125 ,  C12Q2563/137

摘要： Provided is a method for modifying a ssRNA at the 3′ end, said method comprising contacting the strand with a ssRNA 2′-O-methyltransferase in the presence of a co-factor, under conditions which allow for the transfer by the ssRNA 2′-O-methyltransferase of a part of the co-factor onto 3′ end of the ssRNA to form a modified ssRNA, wherein the ssRNA bears 2′-OH group at 3′ terminal nucleotide and wherein the part of the co-factor transferred comprises a reporter group or a functional group.

公开(公告)号：US09988673B2

公开(公告)日：2018-06-05

申请号：US15072677

申请日：2016-03-17

申请人： Vilnius University

发明人： Saulius Klimasauskas ,  Zdislav Stasevskij ,  Edita Kriukiene

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/6853 ,  C12P19/34 ,  C12Q1/6846 ,  C12Q1/6869 ,  C12Q1/6876 ,  C12Q2521/125 ,  C12Q2525/117 ,  C12Q2525/155 ,  C12Q2537/1373

摘要： A method for producing a nucleic acid molecule from a template nucleic acid sequence and a linking unit attached to a primer, which method comprises a step of contacting the template nucleic acid sequence with a nucleic acid polymerase under conditions which allow the nucleic acid polymerase to produce the nucleic acid molecule from the primer based on the template nucleic acid sequence, wherein the linking unit is attached to a target site in the template nucleic acid sequence with a covalent linkage.

公开(公告)号：US20150050699A1

公开(公告)日：2015-02-19

申请号：US14385857

申请日：2013-03-20

申请人： VILNIUS UNIVERSITY

发明人： Virginijus Siksnys ,  Giedrius Gasiunas ,  Tautvydad Karvelis ,  Arvydas Lubys ,  Lolita Zaliauskiene ,  Monika Glemzaite ,  Anja Smith

IPC分类号： C12N15/90 ,  C12N9/22 ,  C12N15/113 ,  C12N15/10

CPC分类号： C12N15/113 ,  C12N9/22 ,  C12N15/102 ,  C12N15/902 ,  C12N15/907 ,  C12N2310/12 ,  C12N2310/14 ,  C12N2310/20 ,  C12N2310/3513 ,  C12N2310/531 ,  C12N2320/00 ,  C12N2800/80 ,  C12P19/34 ,  C12Q1/6811 ,  C12Q2521/301 ,  C12Q2522/101 ,  C12Q2525/121 ,  C12Q2525/151 ,  C12Q2525/301

摘要： Isolation or in vitro assembly of the Cas9-crRNA complex of the Streptococcus thermophilus CRISPR3/Cas system and use for cleavage of DNA bearing a nucleotide sequence complementary to the crRNA and a proto-spacer adjacent motif. Methods for site-specific modification of a target DNA molecule in vitro or in vivo using an RNA-guided DNA endonuclease comprising RNA sequences and at least one of an RuvC active site motif and an HNH active site motif; for conversion of Cas9 polypeptide into a nickase cleaving one strand of double-stranded DNA by inactivating one of the active sites (RuvC or HNH) in the polypeptide by at least one point mutation; for assembly of active polypeptide-polyribonucleotides complex in vivo or in vitro; and for re-programming a Cas9-crRNA complex specificity in vitro and using a cassette containing a single repeat-spacer-repeat unit.

摘要翻译： 分离或体外装配嗜热链球菌CRISPR3 / Cas系统的Cas9-crRNA复合物，并用于裂解含有与crRNA互补的核苷酸序列的DNA和原间隔物相邻基序。 使用包含RNA序列和至少一个RuvC活性位点基序和HNH活性位点基序的RNA引导的DNA核酸内切酶在体外或体内对靶DNA分子进行位点特异性修饰的方法; 用于通过使多肽中的一个活性位点（RuvC或HNH）灭活至少一个点突变，将Cas9多肽转化成切割酶，从而切割一条双链DNA; 用于在体内或体外组装活性多肽 - 多核苷酸复合物; 以及用于在体外重新编程Cas9-crRNA复合物特异性并使用含有单个重复 - 间隔 - 重复单元的盒。

公开(公告)号：US12123031B2

公开(公告)日：2024-10-22

申请号：US16618238

申请日：2018-05-31

申请人： VILNIUS UNIVERSITY

发明人： Virginijus Šikšnys ,  Migle Kazlauskiene ,  Georgij Kostiuk ,  Gintautas Tamulaitis

IPC分类号： C12N9/22 ,  C12N9/12 ,  C12P19/34 ,  C12Q1/6811

CPC分类号： C12N9/22 ,  C12N9/1241 ,  C12P19/34 ,  C12Q1/6811 ,  C12Y207/07 ,  C12N2310/20

摘要： A method of synthetizing cyclic oligoadenylates using a novel catalytic activity of a protein possessing the Palm domain, such as the Cas10 protein, and using such compounds for activation of proteins possessing the CARF doman, such as the Csm6 protein.

公开(公告)号：US20240318354A1

公开(公告)日：2024-09-26

申请号：US18715381

申请日：2022-12-01

申请人： VILNIUS UNIVERSITY

发明人： Linas MAZUTIS ,  Greta LEONAVICIENE ,  Denis BARONAS

IPC分类号： C40B70/00 ,  C12Q1/6806 ,  C40B50/00

CPC分类号： C40B70/00 ,  C12Q1/6806 ,  C40B50/00

摘要： The present invention concerns a method for sequentially attaching a plurality of oligonucleotides to nucleic acids to produce barcoded nucleic acids, wherein the nucleic acids are obtained from a plurality of cells, wherein the plurality of cells are in a plurality of microcapsules, each microcapsule comprising a semi-permeable shell and a core.