公开(公告)号：US10995361B2

公开(公告)日：2021-05-04

申请号：US15876347

申请日：2018-01-22

Applicant: Massachusetts Institute of Technology

Inventor： Fei Chen ,  Asmamaw T. Wassie ,  Shahar Alon ,  Adam Henry Marblestone ,  Anubhav Sinha ,  Andrew Payne ,  Edward Stuart Boyden

IPC: C12Q1/68 ,  C12Q1/6806 ,  C12Q1/6841 ,  C12Q1/6811 ,  C12Q1/682 ,  C12Q1/6855

Abstract: The present invention relates to a method for amplifying at least one target RNA in a fixed and, optionally, expanded biological sample. In an embodiment of the invention, the method comprises incubating the fixed biological sample with a pair of polynucleotides complementary to non-overlapping and proximal sequences of a target RNA, wherein the polynucleotide pair hybridizes to the target RNA; ligating the polynucleotide pair using a ligase; and amplifying the ligation product. The invention further provides methods for detecting and optionally quantifying and/or sequencing the amplification product. As the method comprises hybridizing polynucleotide pairs to a target RNA in a fixed biological sample, the target RNA can be hybridized in situ.

公开(公告)号：US10526649B2

公开(公告)日：2020-01-07

申请号：US15789419

申请日：2017-10-20

Applicant: Massachusetts Institute of Technology ,  President and Fellows of Harvard College

Inventor： Fei Chen ,  Shahar Alon ,  Andrew Payne ,  Asmamaw Wassie ,  Daniel Goodwin ,  Edward Stuart Boyden ,  Evan Daugharthy ,  Jonathan Scheiman

IPC: C12Q1/6874 ,  C12Q1/6841

Abstract: The invention provides in situ nucleic acid sequencing to be conducted in biological specimens that have been physically expanded. The invention leverages the techniques for expansion microscopy (ExM) to provide new methods for in situ sequencing of nucleic acids in a process referred to herein as “expansion sequencing” (ExSEQ).

公开(公告)号：US20180119219A1

公开(公告)日：2018-05-03

申请号：US15789419

申请日：2017-10-20

Applicant: Massachusetts Institute of Technology

Inventor： Fei Chen ,  Shahar Alon ,  Andrew Payne ,  Asmamaw Wassie ,  Daniel Goodwin ,  Edward Stuart Boyden

IPC: C12Q1/6874

CPC classification number: C12Q1/6874 ,  C12Q1/6841 ,  C12Q2545/113

Abstract: The invention provides in situ nucleic acid sequencing to be conducted in biological specimens that have been physically expanded. The invention leverages the techniques for expansion microscopy (ExM) to provide new methods for in situ sequencing of nucleic acids in a process referred to herein as “expansion sequencing” (ExSEQ).

公开(公告)号：US20180216161A1

公开(公告)日：2018-08-02

申请号：US15876347

申请日：2018-01-22

Applicant: Massachusetts Institute of Technology

Inventor： Fei Chen ,  Asmamaw T. Wassie ,  Shahar Alon ,  Adam Henry Marblestone ,  Anubhav Sinha ,  Andrew Payne ,  Edward Stuart Boyden

IPC: C12Q1/6806 ,  C12Q1/6811 ,  C12Q1/682 ,  C12Q1/6855

CPC classification number: C12Q1/6806 ,  C12Q1/6811 ,  C12Q1/682 ,  C12Q1/6841 ,  C12Q1/6855 ,  C12Q2600/158 ,  C12Q2600/16 ,  C12Q2600/166 ,  C12Q2533/107 ,  C12Q2543/101

Abstract: The present invention relates to a method for amplifying at least one target RNA in a fixed and, optionally, expanded biological sample. In an embodiment of the invention, the method comprises incubating the fixed biological sample with a pair of polynucleotides complementary to non-overlapping and proximal sequences of a target RNA, wherein the polynucleotide pair hybridizes to the target RNA; ligating the polynucleotide pair using a ligase; and amplifying the ligation product. The invention further provides methods for detecting and optionally quantifying and/or sequencing the amplification product. As the method comprises hybridizing polynucleotide pairs to a target RNA in a fixed biological sample, the target RNA can be hybridized in situ.