公开(公告)号：US20200261879A1

公开(公告)日：2020-08-20

申请号：US16649068

申请日：2018-09-28

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  Makiko N. Hatori ,  Leqian Liu ,  Samuel Kim ,  Cyrus Modavi

IPC: B01J13/08 ,  C12Q1/6869 ,  C12Q1/6844 ,  B01F17/00

Abstract: The methods described herein, referred to as particle-templated emulsification (PTE), provide an improved approach for generating a monodisperse emulsion that encapsulates target particles of interest without requiring the use of a microfluidic device. Monodisperse droplets may be effectively obtained by using monodisperse particles to template the formation of droplets, which can include, e.g., monodisperse single-emulsion droplets, multiple-emulsion droplets, or Giant Unilamellar Vesicles (GUV), without destroying the integrity of the droplets.

公开(公告)号：US20230227901A1

公开(公告)日：2023-07-20

申请号：US18010188

申请日：2021-06-24

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  Iain Clark ,  Peng Xu ,  Cyrus Modavi

IPC: C12Q1/6869 ,  B01L3/00

CPC classification number: C12Q1/6869 ,  B01L3/502792 ,  C12Q2600/156 ,  C12Q2600/16 ,  B01L2300/0867 ,  B01L2300/0645 ,  B01L2300/0654 ,  B01L2200/0652 ,  B01L2400/0622 ,  B01L2200/16 ,  B01L2400/0427

Abstract: Methods for selectively adding one or more reagents are provided. In certain aspects, the methods include selectively merging one or more droplets of a plurality of droplets with one or more droplets of a plurality of reagent droplets based on detection of a property. Systems, devices and kits for practicing the subject methods are also provided. The subject disclosure may find use in a wide variety of applications, such as increasing the accuracy and/or efficiency of single-cell sequencing, detection of cancer or other diseases, monitoring disease progression, analyzing the DNA or RNA content of cells, and other applications in which it is desired to detect and/or quantify specific target cells.

公开(公告)号：US10829798B2

公开(公告)日：2020-11-10

申请号：US15514777

申请日：2015-09-29

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA ,  David J. Sukovich ,  Cyrus Modavi

Inventor： John Christopher Anderson ,  David J. Sukovich ,  Cyrus Modavi ,  Markus De Raad

IPC: C12P19/34 ,  C12Q1/48 ,  G01N33/542

Abstract: Traditional enzyme characterization methods are low-throughput, and therefore limit engineering efforts in synthetic biology and biotechnology. Here we propose a DNA-linked enzyme-coupled assay (DLEnCA) to monitor enzyme reactions in a high-throughput manner. Throughput is improved by removing the need for protein purification and by limiting the need for liquid chromatography mass spectrometry (LCMS) product detection by linking enzymatic function to DNA modification. DLEnCA is generalizable for many enzymatic reactions, and here we adapt it for glucosyltransferases, methyltransferases, and oxidoreductases. The assay utilizes cell free transcription/translation systems to produce enzymes of interest, while UDP-Glucose and T4-β-glucosyltransferase are used to modify DNA, which is detected post-reaction using qPCR or similar means of DNA analysis. For monitoring methyltransferases, consumption of SAM is observed by coupling to EcoRI methyltransferase. For monitoring oxidoreductases, consumption of NADH is observed by coupling to Taq or E. coli DNA ligase. OleD and two glucosyltransferases from Arabidopsis were used to verify the assay's generality toward glucosyltransferases. Two methyltransferases from human and Arabidopsis were used to verify the assay's generality towards methyltransferases. We show DLEnCA's utility by mapping out the substrate specificity for these enzymes and observing the multiple steps of a biosynthetic pathway.