Patent applications filed by the Penn State Research Foundation
The present PCT and United States patent applications relate to the
transformation of the fruit body tissue of a filamentous fungus
with Agrobacterium. The genetically modified fungi can serve as
biofermentators for the mass production of commercial products. As examples of
filamentous fungi, the applicant mentions fungi belonging to phyla Ascomycota
and Basidiomycota such as Coprinus, Agaricus,
Morchella , and Coriolus, among others. In particular, the
invention contemplates the transformation of the cultivated mushroom
Agaricus bisporus, which accounts for 38% of the world production of
cultivated mushrooms.
Specific Patent Information
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Patent Number
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Specific Patent Information
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Assignee
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WO
2002/00896 A2
- Earliest priority - 28 June 2000
- Filed - 28 June 2001
- OPI - 3 January 2002
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Title - Methods and compositions for highly effective
transformation of filamentous fungi
| Claim 1
A method of transforming filamentous fungi comprising:
introducing to a fruit body tissue cell of said fungi a polynucleotide
construct said construct comprising sequences for the expression of a structural
gene or for the inhibition of an endogenous gene, the presence of which is
desired in said fungi cell.
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| Claim 16
A filamentous fungi transformation method comprising:
A) obtaining an Agrobacterium vector, said vector comprising a
polynucleotide sequence the presence of which is desired in a recipient
filamentous fungi cell; and
B) introducing said vector to fruit body tissue
cells of said fungi in the presence of an active vir
Agrobacterium region and a vir inducing compound.
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| Claim 26
A method for transforming a fungi cell comprising:
A) obtaining an Agrobacterium-derived Ti plasmid, said plasmid
comprising a polynucleotide sequence the expression of which is desired in a
host fungal cell;
B) introducing said plasmid to said cell in the presence
of vir active genes and a vir inducing agent.
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| Claim 30
A method for transformation of Agaricus bisporus cells comprising:
A) obtaining an Agrobacterium vector, said vector comprising a
polynucleotide sequence the presence of which is desired in a recipient
filamentous fungi cell; and
B) introducing said vector to fruit body tissue
cells of said Agaricus bisporus cells in the presence of an active
vir Agrobacterium region and a vir inducing compound.
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The claims as filed in the PCT application recite:
- a method for transforming any filamentous fungus by
introducing into the fruit body tissue a construct for the
expression of a structural gene or for the inhibition of an endogenous gene; and
- a method for transforming the fruit body tissue of
any filamentous fungus and Agaricus
bisporus in particular with an Agrobacterium vector in
the presence of a vir acting region and a vir inducing
compound.
Although these claims are fairly broad as they refer to the transformation of
any filamentous fungus without specifying a
method in one case and specifying Agrobacterium in another claim, an
apparent limitation lies in the type of fungal tissue to be transformed. The
invention recites only transformation of fruit body
tissue. It remains to be seen whether granted claims will have the same
breadth as the filed claims.
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Penn State Research Foundation
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US
6964866
- Earliest priority - 28 June 2000
- Filed - 28 June 2001
- Granted - 15 November 2005
- Expected expiry - 1 September 2022 (patent term adjustment: 431 days)
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Title - Methods and compositions for highly efficient
transformation of filamentous fungi
Claim 1
A method of transforming Agaricus bisporus comprising:
(a) introducing to fruit body tissue cells of said Agaricus bisporus
a polynucleotide construct desired in said Agaricus bisporus, said
construct comprising a promoter sequence capable of regulating expression in
Agaricus bisporus cells, wherein said introducing is by
Agrobacterium-mediated transformation by co-cultivation of vir
induced Agrobacterium with Agaricus fruit body tissue cells;
and
(b) thereafter regenerating a transgenic Agaricus bisporus
from said fruit body tissue cells.
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Claim 7
An Agaricus bisporus transformation method comprising:
(a) obtaining an Agrobacterium vector, said vector comprising a
polynucleotide sequence the presence of which is desired in a recipient
Agaricus bisporus cell, said polynucleotide sequence operatively linked
to a promoter sequence capable of regulating expression in Agaricus
bisporus cells;
(b) introducing said vector to fruit body tissue cells
of said Agaricus bisporus in the presence of an active vir
Agrobacterium region and a vir inducing compound by
co-cultivation of vir induced Agrobacterium with
Agaricus fruit body tissue cells; and
(c) regenerating a
transgenic Agaricus bisporus from said fruit body cells.
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Claim 14
A method for transformation of Agaricus bisporus fruit body tissue
cells comprising:
(a) obtaining an Agrobacterium vector, said vector comprising a
polynucleotide sequence the presence of which is desired in a recipient
Agaricus bisporus fruit body tissue cell, said polynucleotide sequence
operatively linked to a promoter sequence capable of regulating expression in
Agaricus bisporus cells and introducing said vector to said
Agaricus bisporus fruit body tissue cells in the presence of an active
vir Agrobacterium region and a vir inducing compound
by co-cultivation of vir induced Agrobacterium with
Agaricus fruit body tissue cells.
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Granted United States patent US 6964866 recites an
Agrobacterium-mediated transformation method of Agaricus
bisporus, where fruit body tissue cells of A. bisporus are
introduced to a vir-induced Agrobacterium.
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US
2005/70007 A1
- Earliest priority - 28 June 2000
- Filed - 30 September 2004
- Application pending
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Title - Methods and compositions for highly efficient
transformation of filamentous fungi
Claim 1
A method of transforming filamentous fungi comprising:
- introducing to a fruit body tissue cells of said fungi a polynucleotide
construct said construct comprising sequences for the expression of a structural
gene or for inhibition of an endogenous gene, the presence of which is desired
in said fungi cell.
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Claim 8
A filamentous fungi transformation method comprising:
(a) obtaining an Agrobacterium vector, said vector comprising a
polynucleotide sequence the presence of which is desired in a recipient
filamentous fungi cell, and
(b) introducing said vector to fruit body
tissue cells of said fungi in the presence of an active vir
Agrobacterium region and a vir inducing compound.
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Claim 10
A method for transforming a fungi cell comprising:
(a) obtaining an Agrobacterium derived ti-plasmid, said plasmid
comprising a polynucleotide sequence the expression of which is desired in a
host fungal cell,
(b) introducing said plasmid to said cell in the presence
of vir active genes and a vir inducing agent.
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United States patent application US 2005/070007 is a
divisional of now granted US 6964866 (see
above).
Independent claim 1 in this application recites a method of transforming
any filamentous fungus by using fruit tissue body
cells that is not limited to
Agrobacterium-mediated transformation.
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| Remarks |
- National phase entry of WO 2002/000896 in Canada (CA 2452183) and Europe (EP
1409693) are pending.
- National phase entry of WO 2002/000896 in Australia (AU 2001271560) has
lapsed on 20 December 2003.
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