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    Methods and systems for cell state quantification
    53.
    发明授权
    Methods and systems for cell state quantification 有权

    公开(公告)号：US10119975B2

    公开(公告)日：2018-11-06

    申请号：US15363243

    申请日：2016-11-29

    Applicant: Ginkgo Bioworks, Inc.

    Inventor： Reshma Shetty ,  Thomas F. Knight, Jr. ,  Randall D. Rettberg

    IPC: C12N15/70 ,  G01N33/68 ,  C12N15/74 ,  C40B40/08

    Abstract: Systems, methods, libraries, kits, and computer software tools are provided for designing and producing engineered cells. Such engineered cells can be used for cell state quantification, such as genome, transcriptome and/or proteome quantification. In one aspect, an engineered cell having a plurality of artificially designed oligonucleotides introduced into the genome of the cell is provided. The oligonucleotides are each located in proximity of a gene of interest encoding a protein of interest, and are different from one another. The oligonucleotides can each encode a unique peptide tag for each protein of interest, wherein each peptide tag has a unique quantitatively measurable value such as mass-to-charge ratio which can be quantified by a mass spectrometer. The engineered cell is capable of expressing a plurality of proteins of interest each fused to its corresponding unique peptide tag, wherein each peptide tag is capable of being released therefrom.

    VARIANT LIBRARIES OF THE IMMUNOLOGICAL SYNAPSE AND SYNTHESIS THEREOF
    57.
    发明申请
    VARIANT LIBRARIES OF THE IMMUNOLOGICAL SYNAPSE AND SYNTHESIS THEREOF 审中-公开

    公开(公告)号：US20180171509A1

    公开(公告)日：2018-06-21

    申请号：US15844395

    申请日：2017-12-15

    Applicant: Twist Bioscience Corporation

    Inventor： Anthony COX ,  Siyuan Chen

    IPC: C40B40/08 ,  G01N33/68 ,  G01N33/50 ,  C12N15/10 ,  A61K39/00

    Abstract: Disclosed herein are methods for the generation of highly accurate nucleic acid libraries encoding for predetermined variants of a nucleic acid sequence. The nucleic acid sequence may encode for all or part of a TCR or a TCR-binding antigen. The degree of variation may be complete, resulting in a saturated variant library, or less than complete, resulting in a non-saturating library of variants. The variant nucleic acid libraries described herein may designed for further processing by transcription or translation. The variant nucleic acid libraries described herein may be designed to generate variant RNA, DNA and/or protein populations. Further provided herein are method for identifying variant species with increased or decreased activities, with applications in regulating biological functions and the design of therapeutics for treatment or reduction of a disease, such as cancer.

    Construction of pool of interfering nucleic acids covering entire RNA target sequence and related compositions
    59.
    发明授权
    Construction of pool of interfering nucleic acids covering entire RNA target sequence and related compositions 有权

    公开(公告)号：US09944928B2

    公开(公告)日：2018-04-17

    申请号：US14791157

    申请日：2015-07-02

    Applicant: York Yuan Yuan Zhu

    Inventor： York Yuan Yuan Zhu

    IPC: C12N15/11 ,  C12N15/113 ,  C40B40/08 ,  C40B50/06

    CPC classification number: C12N15/113 ,  C12N15/111 ,  C12N2310/14 ,  C12N2330/31 ,  C40B40/08 ,  C40B50/06

    Abstract: The present invention provides a PCR based high-throughput method for preparing full-sites siRNA polynucleotide pool, comprising: DNase I random digestion; Loop-1 phosphate linker ligation; single PCR amplification; a type III restriction/modification enzyme digestion; blunt ending; Loop-2 phosphate linker ligation; double primer PCR; FokI digestion and cloning into an siRNA expression vector. The present invention enables the use of a type III restriction/modification enzyme linkers mediated PCR method for high-throughput preparing an siRINA polynucleotide pool, in which the functional length of siRNAs can be controllably distributed from 19-23 bp, thus completely mimic the natural siRNA length diversity, specially suitable for RNAi therapeutic targets screening. The present invention overcomes the bottlenecks and drawbacks of conventional siRNA polynucleotide pool construction technologies.

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