Xiaochun C Fan

US Patent:

20200190540, Jun 18, 2020

Filed:

Oct 30, 2019

Appl. No.:

16/668528

Inventors:

- Wilmington DE, US
Xiaochun Fan - West Chester PA, US

International Classification:

C12N 15/90
C12N 15/113
C12N 9/16
C12N 15/10

Abstract:

Non-conventional yeasts are disclosed herein comprising at least one RNA-guided endonuclease (RGEN) comprising at least one RNA component that does not have a 5′-cap. This uncapped RNA component comprises a sequence complementary to a target site sequence in a chromosome or episome in the yeast. The RGEN can bind to, and optionally cleave, one or both DNA strands at the target site sequence. An example of an RGEN herein is a complex of a Cas9 protein with a guide RNA. A ribozyme is used in certain embodiments to provide an RNA component lacking a 5′-cap. Further disclosed are methods of genetic targeting in non-conventional yeast.

US Patent:

20200157516, May 21, 2020

Filed:

Jan 27, 2020

Appl. No.:

16/752887

Inventors:

- WILMINGTON DE, US
XIAOCHUN FAN - WEST CHESTER PA, US
ETHEL NOLAND JACKSON - GREENVILLE DE, US

International Classification:

C12N 9/22
C12N 15/90
C12N 15/86
C12N 15/62
C12N 15/10
C12N 9/96

Abstract:

A composition is disclosed that comprises at least one protein component of an RNA-guided endonuclease (RGEN) and at least one cell-penetrating peptide (CPP), wherein the RGEN protein component and CPP are covalently or non-covalently linked to each other in an RGEN protein-CPP complex. The RGEN protein-CPP complex can traverse (i) a cell membrane, or (ii) a cell wall and cell membrane, of a cell. The RGEN protein component of an RGEN protein-CPP complex in certain embodiments can be associated with a suitable RNA component to provide an RGEN capable of specific DNA targeting. Further disclosed are compositions comprising at least one protein component of a guide polynucleotide/Cas endonuclease complex and at least one CPP, as well as methods of delivering RGEN proteins into microbial cells, as well as methods of targeting DNA with RGENs.

US Patent:

20190300877, Oct 3, 2019

Filed:

Apr 9, 2019

Appl. No.:

16/378955

Inventors:

- Palo Alto CA, US
XIAOCHUN FAN - WEST CHESTER PA, US
RYAN L. FRISCH - NEWARK DE, US

International Classification:

C12N 15/11
C12N 15/81
C12N 15/90
C12N 15/113
C12N 9/22

Abstract:

Compositions and methods are provided for editing nucleotides and/or altering target sites in the genome of a cell. The methods and compositions employ a recombinant DNA construct comprising a tRNA promoter operably linked to a polynucleotide encoding a single guide RNA, wherein said recombinant DNA construct does not comprise a nucleotide sequence encoding a ribozyme, wherein said guide RNA is capable of forming a guide RNA/Cas endonuclease complex, wherein said complex can bind to and cleave a target site sequence in the genome of a cell such as a microbial cell. The present disclosure further describes methods and compositions employing a recombinant DNA construct comprising a tRNA promoter operably linked to a spacer sequence and a polynucleotide encoding a single guide RNA, wherein said recombinant DNA construct does not comprise a nucleotide sequence encoding a ribozyme, wherein said guide RNA is capable of forming a guide RNA/Cas endonuclease complex, wherein said complex can bind to and cleave a target site sequence in the genome of a non-conventional yeast.

US Patent:

20190144897, May 16, 2019

Filed:

Jul 20, 2016

Appl. No.:

15/746094

Inventors:

- Wilmington DE, US
Xiaochun Fan - West Chester PA, US
Dongming Xie - Newark DE, US
Himanshu H. Dhamankar - Greenville DE, US

International Classification:

C12P 7/64
C12N 9/00

Abstract:

Recombinant microbial cells comprising an engineered LCDA production pathway that comprises at least one up-regulated long-chain acyl-CoA synthetase (ACoS) are disclosed. These recombinant microbial cells are capable of producing one or more long-chain dicarboxylic acid (LCDA) products from a long-chain fatty acid-comprising substrate. Methods of using recombinant microbial cells to produce LCDAs are also disclosed.

US Patent:

20190100737, Apr 4, 2019

Filed:

Dec 13, 2018

Appl. No.:

16/218808

Inventors:

- Wilmington DE, US
XIAOCHUN FAN - WEST CHESTER PA, US
ETHEL NOLAND JACKSON - GREENVILLE DE, US

International Classification:

C12N 9/22
C12N 15/90
C12N 9/96
C12N 15/62
C12N 15/86
C12N 15/10

Abstract:

A composition is disclosed that comprises at least one protein component of an RNA-guided endonuclease (RGEN) and at least one cell-penetrating peptide (CPP), wherein the RGEN protein component and CPP are covalently or non-covalently linked to each other in an RGEN protein-CPP complex. The RGEN protein-CPP complex can traverse (i) a cell membrane, or (ii) a cell wall and cell membrane, of a cell. The RGEN protein component of an RGEN protein-CPP complex in certain embodiments can be associated with a suitable RNA component to provide an RGEN capable of specific DNA targeting. Further disclosed are compositions comprising at least one protein component of a guide polynucleotide/Cas endonuclease complex and at least one CPP, as well as methods of delivering RGEN proteins into microbial cells, as well as methods of targeting DNA with RGENs.

US Patent:

20170335300, Nov 23, 2017

Filed:

Nov 3, 2015

Appl. No.:

15/523741

Inventors:

- Wilmington DE, US
Xiaochun Fan - West Chester PA, US
Ethel Noland Jackson - Greenville DE, US

International Classification:

C12N 9/22
C12N 15/90
C12N 15/10
C12N 15/86
C12N 15/62
C12N 9/96

Abstract:

A composition is disclosed that comprises at least one protein component of an RNA-guided endonuclease (RGEN) and at least one cell-penetrating peptide (CPP), wherein the RGEN protein component and CPP are covalently or non-covalently linked to each other in an RGEN protein-CPP complex. The RGEN protein-CPP complex can traverse (i) a cell membrane, or (ii) a cell wall and cell membrane, of a cell. The RGEN protein component of an RGEN protein-CPP complex in certain embodiments can be associated with a suitable RNA component to provide an RGEN capable of specific DNA targeting. Further disclosed are compositions comprising at least one protein component of a guide polynucleotide/Cas endonuclease complex and at least one CPP, as well as methods of delivering RGEN proteins into microbial cells, as well as methods of targeting DNA with RGENs.

US Patent:

20170226533, Aug 10, 2017

Filed:

Jul 5, 2015

Appl. No.:

15/503383

Inventors:

- Wilmington DE, US
Xiaochun FAN - West Chester PA, US

Assignee:

E I DU PONT DE NEMOURS AND COMPANY - Wilmington DE

International Classification:

C12N 15/90

Abstract:

Non-conventional yeasts are disclosed herein comprising at least one RNA-guided endonuclease (RGEN) comprising at least one RNA component that does not have a 5′-cap. This uncapped RNA component comprises a sequence complementary to a target site sequence in a chromosome or episome in the yeast. The RGEN can bind to, and optionally cleave, one or both DNA strands at the target site sequence. An example of an RGEN herein is a complex of a Cas9 protein with a guide RNA. A ribozyme is used in certain embodiments to provide an RNA component lacking a 5′-cap. Further disclosed are methods of genetic targeting in non-conventional yeast.

US Patent:

20140186906, Jul 3, 2014

Filed:

Dec 18, 2013

Appl. No.:

14/132022

Inventors:

- Wilmington DE, US
Pamela L. Sharpe - Wilmington DE, US
Narendra S. Yadav - Wilmington DE, US
Xiaochun Fan - Wilmington DE, US
Michael Dauner - Wilmington DE, US

Assignee:

E I DU PONT DE NEMOURS AND COMPANY - Wilmington DE

International Classification:

C12P 7/64

US Classification:

435134, 4352542

Abstract:

Recombinant microbial cells are disclosed herein that produce an oil comprising at least 28 percent eicosapentaenoic acid (EPA) measured as a weight percent of dry cell weight. These cells may comprise a polynucleotide sequence encoding an active acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT) comprising at least one amino acid mutation in a membrane-bound O-acyltransferase motif. In addition, the cells may comprise a down-regulation of an endogenous polynucleotide sequence encoding Sou2 sorbitol utilization protein, and/or one or more polynucleotides encoding phospholipid:diacylglycerol acyltransferase (PDAT), delta-12 desaturase, a dihomo-gamma-linolenic acid (DGLA) synthase multizyme, delta-8 desaturase, malonyl-CoA synthetase (MCS), or acyl-CoA:lysophosphatidic acid acyltransferase (LPAAT). Also disclosed are methods of using the recombinant microbial cells to produce oil containing omega-3 polyunsaturated fatty acids such as EPA.