論文の残滓

特許実務に関するあれこれ。何かあれば→ imperatorkm@じーめーる.com

欧州における動植物関連出願の審査・審理停止

2017年に改正されたRule 28(2) EPCについては、以前アップしたように、Article 53(b)の解釈に関するブロッコリ事件・トマト事件における拡大審判部の判断と抵触するとの判断が審判部から示されました。

 

nannosono.hatenablog.com

欧州特許条約上、規則が法令に抵触すると法令が優先されるため(Article 164(2))、規則は実質無効化されます。

また、一般的な育種方法、いわゆる、交雑・選抜を繰り返す方法で生み出される動植物の特許出願の審査及び審理においては、Article 53(b)+Rule28 (2)がかなり大きな問題となっているため、規則が無効化されたことで、審査・審理が止まるであろうと想定されていました。

そして、ついにEPOから正式に審査・審理の停止がアナウンスされました。

 

www.epo.org

審査・審理の停止は、EPOの長官から拡大審判部に対して、Article 164(2)に基づいて、Rule28 (2)がArticle 53(b)と整合性がとれているかとの質問に対して,拡大審判部の回答が得られるまででであるため、おおよそ2-3年は続くことになると想定されます。

EPOの審査は遅いので、今、EPOにpendingしている案件については、下手をすると、審査・審理のけりが付く頃には特許権の存続期間はほとんど残ってないかもしれませんね…笑

 

なお、サーチに関しては引き続き行われますので、拡張SRは通常どおり発行される考えられます。

 また、クレームがGMの動植物に関することが明確である場合、そもそもArticle 53(b)+Rule28 (2)の規定とは関係ないため(審査ガイドライン G-II 5.4 Plant and animal varieties, essentially biological processes for the production of plants or animals、特に、5.4.2.1参照)、審査・審理は停止しないことになります。

JP Patent status and claim 1 of Broad's CRISPR-Cas9 Patents.

Broadの日本特許3件

 

特許第6203879号

【請求項1】
クラスター化等間隔短鎖回分リピート(CRISPR)-CRISPR関連(Cas)(CRISPR-Cas)ベクター系であって、
I. CRISPR-Cas系キメラRNA(chiRNA)ポリヌクレオチド配列をコードするヌクレオチド配列に作動可能に結合している第1の調節エレメントであって、
前記ポリヌクレオチド配列が、
(a)真核細胞中の標的配列にハイブリダイズする、10~30ヌクレオチドの長さを有するガイド配列、
(b)トランス活性化CRISPR RNA(tracr)メイト配列、及び
(c)tracrRNA配列
を含み、
(a)、(b)及び(c)が、5’から3’配向で配置されており、
前記tracrRNA配列が、50以上のヌクレオチドの長さを有する、
第1の調節エレメントと、
II. 真核細胞の核中の検出可能な量のII型Cas9タンパク質の蓄積をドライブするために十分な強度の、1つ以上の核局在化配列を含む前記Cas9タンパク質をコードするヌクレオチド配列に作動可能に結合している第2の調節エレメントと
を含む1つ以上のベクターを含み;
成分I及びIIは、前記系の同じ又は異なるベクター上に位置し;
前記ヌクレオチド配列が転写されると:
前記chiRNAは、前記II型Cas9タンパク質へと集合し、前記II型Cas9タンパク質と複合体を形成し、
前記tracrメイト配列は、前記tracrRNA配列にハイブリダイズし、
前記ガイド配列は、前記真核細胞中の前記標的配列への配列特異的結合を指向し、
それによって、(1)前記真核細胞中の前記標的配列にハイブリダイズされる前記ガイド配列、及び(2)前記tracrRNA配列にハイブリダイズされる前記tracrメイト配列と複合体形成している前記II型Cas9タンパク質を含むCRISPR複合体が形成される、
CRISPR-Casベクター系。

 

・II型CRISPR-Cas9

・ハイブリダイズの長さ限定

・tracrRNAの長さ限定

特許第6420273号
【請求項1】 エンジニアリングされた、天然に存在しないクラスター化等間隔短鎖回分リピート(CRISPR)-CRISPR関連(Cas)(CRISPR-Cas)ベクター系であって、 a)ガイド配列、tracrRNA及びtracrメイト配列を含む1つ以上のCRISPR-Cas系ポリヌクレオチド配列をコードする1つ以上のヌクレオチド配列に作動可能に結合している第1の調節エレメントであって、前記ガイド配列は、真核細胞中のポリヌクレオチド遺伝子座中の1つ以上の標的配列にハイブリダイズする、第1の調節エレメント、 b)II型Cas9タンパク質をコードするヌクレオチド配列に作動可能に結合している第2の調節エレメント を含む1つ以上のベクターを含み、 成分(a)及び(b)は、前記系の同じ又は異なるベクター上に位置し、 前記CRISPR-Cas系は、前記Cas9タンパク質をコードする前記ヌクレオチド配列とともに発現される2つ以上の核局在化シグナル(NLS)を含み、少なくとも1つのNLSが、前記Cas9タンパク質のアミノ末端に若しくはその付近に存在し、及び、少なくとも1つのNLSが、前記Cas9タンパク質のカルボキシ末端に若しくはその付近に存在し、 それによって、前記1つ以上のガイド配列が、真核細胞中の前記1つ以上のポリヌクレオチド遺伝子座を標的とし、前記Cas9タンパク質が、前記1つ以上のポリヌクレオチド遺伝子座を開裂し、それによって、前記1つ以上のポリヌクレオチド遺伝子座の配列が、改変される、 CRISPR-Casベクター系。

・II型CRISPR-Cas9
・NLSが2つ以上

・N末端、C末端にNLS一個ずつ配置

 

特許第6395765号

CRISPR複合体成分を含む組成物であって、
前記CRISPR複合体成分が、
I. CRISPR-Cas系ポリヌクレオチド配列(複数の場合も有り)であって、
(a)RNAを含み、ポリヌクレオチド遺伝子座中の標的配列にハイブリダイズし得る、エンジニアリングされたガイド配列、
(b)RNAを含むtracrメイト配列、及び
(c)RNAを含むtracrRNA配列
を含み、
(a)、(b)及び(c)が、5’~3’配向で配置されている、
CRISPR-Cas系ポリヌクレオチド配列、
II. II型Cas9タンパク質
を含み、
前記tracrメイト配列が、前記tracrRNA配列にハイブリダイズし、前記ガイド配列が、標的配列へのCRISPR複合体の配列特異的結合を指向し、
前記CRISPR複合体が、前記II型Cas9タンパク質であって、(1)前記標的配列にハイブリダイズされる前記ガイド配列及び(2)前記tracrRNA配列にハイブリダイズされる前記tracrメイト配列と複合体形成している、前記II型Cas9タンパク質を含み、
前記II型Cas9タンパク質が、黄色ブドウ球菌(Staphylococcus aureus)Cas9(SaCas9)である又はこれを含む、

 

・小型Cas9(SaCas9)

EP Patent status and claim 1 of UC Berkeley‘s CRISPR-Cas9 Patents.

執筆用メモ

 

UCBの欧州特許

一本鎖型については本命、欧州では二本鎖型を狙った出願を既に権利化済。

 

EP2800811B:口頭審理未定(補正クレーム提出済(2018/10/25))

1.  A method of modifying a target DNA, the method comprising contacting the target DNA with a complex comprising:

  (a) a Cas9 polypeptide and

  (b) a single-molecule DNA-targeting RNA comprising:

    (i) a DNA-targeting segment comprising a nucleotide sequence that is complementary to a sequence in the target DNA, and

    (ii) a protein-binding segment that interacts with said Cas9 polypeptide, wherein the protein-binding segment comprises two complementary stretches of nucleotides that hybridize to form a double stranded RNA (dsRNA) duplex,

   wherein said two complementary stretches of nucleotides are covalently linked by intervening nucleotides,

   wherein said contacting is in vitro or in a cell ex vivo; and

   wherein said modifying is cleavage of the target DNA.

 

・一本鎖型の基本特許

 

EP3241902B:異議申立あり

1.  A composition comprising:

  (a) a chimeric Cas9 protein, or a polynucleotide encoding said chimeric Cas9 protein,

     wherein the chimeric Cas9 protein comprises a modified Cas9 protein having reduced nuclease activity compared to the corresponding wild-type Cas9, and comprises a heterologous polypeptide that:

    (i) has DNA modifying activity, or

    (ii) exhibits the ability to increase or decrease transcription, or

    (iii) has enzymatic activity that modifies a polypeptide associated with DNA; and

  (b) a DNA-targeting RNA, or one or more DNA polynucleotides encoding said DNA-targeting RNA, wherein said DNA-targeting RNA comprises:

    (i) a DNA-targeting segment comprising a nucleotide sequence that is complementary to a sequence in a target DNA, and

    (ii) a protein-binding segment that interacts with said chimeric Cas9 protein,

    wherein the protein-binding segment comprises two complementary stretches of nucleotides that hybridize to form a double stranded RNA (dsRNA) duplex.

 

・ニッカーゼ型Cas9およびdCas9をカバーする形態。

・ガイド鎖は、一本鎖型に限定されず、二本鎖型を含む。

神大 西田先生のバイオパレットは、sgRNA+dCas9-AIDを使用しているはずなので、(a)(i)で引っかかってる可能性。

EP Patent status and claim 1 of VILNIUS‘s CRISPR-Cas9 Patents.

執筆用メモ

 

VILNIUS大学の欧州特許、OA苦戦中であったが、1月23日にGrantがでようで。

5者の中で最先でFileしているので、代理人がもっと優秀であれば、結果は違ったかも知れません。明日は我が身…。

 

EP2828386

1 . A in-vitro method for the site-specific modification of a target DNA molecule, the method comprising

     contacting, under suitable conditions,
         a target DNA molecule; and
         an RNA-guided DNA endonuclease comprising at least one RNA sequence and at least one of an RuvC active site motif and an HNH active site motif;

       wherein

       the RNA-guided DNA endonuclease is a Cas9-crRNA complex

       said Cas9-crRNA complex comprising a Cas9, crRNA and tracrRNA
       to result in the target DNA molecule modified in a region that is determined by the complimentary binding of the crRNA sequence to the target DNA molecule,

      the method further comprising assembling the popeptide-polyribonucleotides complex in-vitro by incubating said components of the complex under conditions suitable for complex assembly,

      wherein the target DNA is double stranded or single stranded and wherein a double stranded target DNA contains a proto-spacer adjacent motif.

 

・日米欧共にin vitro assemblyが必須なので、特許としては…

EP Patent status and claim 1 of Toolgen‘s CRISPR-Cas9 Patents.

執筆用メモ

 

ツールジェンの欧州特許(2019/02/22)

真核生物のデータを含めて初めてFileしたのはToolgen。

ただし、Sienceの論文公開後であり、全般的に進歩性?

ガイド鎖末端のGG付加がポイントか。

 

EP2912175:異議申立中(口頭審理未定)

1.  A Type II Clustered Regularly interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system for introducing double-stranded breaks into a target DNA sequence in a mammalian cell,

   wherein the Type II CRISPR/Cas9 system comprises:

       a. a Cas9 protein with one nuclear localization signal (NLS),

       wherein the NLS is at the C-terminus, or a nucleic acid encoding the Cas9 protein; and

       b. a single-chain guide RNA comprising a CRISPR RNA (crRNA) portion fused to a trans-activating crRNA (tracrRNA) portion.

 

・Cas9に1個NLSが付加された、sgRNAを利用するCRISPR-Cas9全般をカバー。NLS1個ついただけで進歩性をクリアというのが謎。

・第二医薬用途クレームの形式であるため、通常は解釈されない用途が解釈されている…?

 

EP Patent status and claim 1 of SIGMA ALDRICH‘s CRISPR-Cas9 Patents.

執筆用メモ
 
SIGMA ALDRICHの欧州特許(2019/02/22)
SIGMAの技術の特徴は、HR修復を使った核酸の導入と、ニッカーゼ型で切る点。
 
EP3138910B:異議申立中(口頭審理未定)

1.  A method for integrating an exogenous sequence into a chromosomal sequence of a eukaryotic cell, the method comprising:

  a) introducing into the eukaryotic cell

      (i) at least one RNA-guided endonuclease comprising at least one nuclear localization signal or nucleic acid encoding at least one RNA-guided endonuclease comprising at least one nuclear localization signal,

         wherein the at least one RNA-guided endonuclease is a clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated (Cas) (CRISPR/Cas) type II system protein and the CRISPR/Cas type II system protein is a Cas9 protein,

      (ii) at least one guide RNA or DNA encoding at least one guide RNA, and

      (iii) at least one donor polynucleotide comprising the exogenous sequence; and

  b) culturing the eukaryotic cell such that the guide RNA guides the RNA-guided endonuclease to a target site in the chromosomal sequence where the RNA-guided endonuclease introduces a double-stranded break, and the double-stranded break is repaired by a DNA repair process such that the exogenous sequence is integrated into the chromosomal sequence,

   wherein the method does not comprise a process for modifying the germ line genetic identity of a human being and,

   wherein the method does not comprise a method for treatment of the human or animal body.

 

・HRの修復を使った核酸の導入

 

EP3138911B:異議申立中(始まったばっかり、口頭審理未定)

1.  A method for modifying a chromosomal sequence in a eukaryotic cell by integrating a donor sequence, the method comprising:

  a) introducing into the eukaryotic cell

      (i) at least one RNA-guided endonuclease comprising at least one nuclear localization signal or nucleic acid encoding at least one RNA-guided endonuclease comprising at least one nuclear localization signal,

       wherein the at least one RNA-guided endonuclease is a clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated (Cas) (CRISPR/Cas) type II system protein and the CRISPR/Cas type II system protein is a Cas9 protein,

      (ii) at least one guide RNA or DNA encoding at least one guide RNA, and

      (iii) a donor polynucleotide comprising the donor sequence; and

  b) culturing the eukaryotic cell such that each guide RNA guides an RNA-guided endonuclease to a target site in the chromosomal sequence, the RNA-guided endonuclease introduces a double-stranded break at the target site, and the double-stranded break is repaired by a DNA repair process such that the chromosomal sequence is modified by insertion or substitution of the donor sequence into the chromosomal sequence,

   wherein the target site in the chromosomal sequence is immediately followed by a protospacer adjacent motif (PAM),

   the method does not comprise a process for modifying the germ line genetic identity of a human being and,

   wherein the method does not comprise a method for treatment of the human or animal body by surgery or therapy.

 

・HRの修復を使った核酸の導入

 

EP3138912B:異議申立中(始まったばっかり、口頭審理未定)

1.  A method for modifying a chromosomal sequence in a eukaryotic cell, the method comprising:

  a) introducing into the eukaryotic cell two RNA-guided nickase systems or nucleic acid encoding said systems, and, optionally, a donor polynucleotide,

     wherein each RNA-guided nickase system comprises

      (i) a RNA-guided endonuclease that is modified to cleave one strand of a double-stranded sequence; and

      (ii) a guide RNA comprising a first region having complementarity to a target site in the chromosomal sequence and a second region that interacts with the RNA-guided endonuclease,

          wherein each target site is immediately followed by a protospacer adjacent motif (PAM), and the target sites of the two RNA-guided endonucleases are on opposite strands of the chromosomal sequence; and

  b) culturing the eukaryotic cell such that the two RNA-guided endonucleases cleave opposite strands of the chromosomal sequence in close enough proximity to introduce a double-stranded break in the chromosomal sequence, and repair of the double-stranded break by a DNA repair process leads to modification of the chromosomal sequence,

   wherein the method does not comprise a process for modifying the germ line genetic identity of a human being and,

   wherein the method does not comprise a method for treatment of the human or animal body by surgery or therapy.

 

・HRの修復を使った核酸の導入×ニッカーゼ型

・ニッカーゼは、Cas9に限定されず、RNAガイドエンドヌクレアーゼ(他のCasを含む)

EP Patent status and claim 1 of Broad’s CRISPR-Cas9 Patents.

‘執筆用メモ

欧州におけるBroad Instituteの特許の状況。~2018年10月初旬までに成立済のものまで。

 

EP2764103:口頭審理待ち(2019/03/04、補正クレーム提出済)

1.  An engineered, non-naturally occurring Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR)-CRISPR associated (Cas) (CRISPR-Cas) vector system comprising one or more vectors comprising:

     a) a first regulatory element operably linked to one or more nucleotide sequences encoding one or more CRISPR-Cas system guide RNAs that hybridize with target sequences in polynucleotide loci in a eukaryotic cell, the guide RNA comprising a guide sequence, a tracr sequence, and a tracr mate sequence,

     b) a second regulatory element operably linked to a nucleotide sequence encoding a Type II Cas9 protein, said protein comprising a nuclear localization signal (NLS);

     wherein components (a) and (b) are located on same or different vectors of the system, wherein the tracr sequence is 30 or more nucleotides in length, and

     whereby the one or more guide RNAs target the polynucleotide loci in a eukaryotic cell and the Cas9 protein cleaves the polynucleotide loci,

     whereby sequence of the polynucleotide loci is modified; and, wherein the Cas9 protein and the one or more guide RNAs do not naturally occur together.

 

EP2771468:特許取消→審判請求

1.  A non-naturally occurring or engineered composition comprising:   a Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR)-CRISPR associated (Cas) (CRISPR-Cas) system chimeric RNA (chiRNA) polynucleotide sequence, wherein the polynucleotide sequence comprises

    (a) a guide sequence of between 10 - 30 nucleotides in length, capable of hybridizing to a target sequence in a eukaryotic cell,

    (b) a tracr mate sequence, and

    (c) a tracrRNA sequence

  wherein (a), (b) and (c) are arranged in a 5' to 3' orientation,

  wherein when transcribed, the tracr mate sequence hybridizes to the tracrRNA sequence and the guide sequence directs sequence-specific binding of a CRISPR complex to the target sequence,

  wherein the CRISPR complex comprises a Type II Cas9 protein complexed with (1) the guide sequence that is hybridized to the target sequence, and (2) the tracr mate sequence that is hybridized to the tracrRNA sequence,

  wherein the tracrRNA sequence is 50 or more nucleotides in length.

 

・優先権無効→D3およびD4に基づく新規性欠如

D3:MALI P. ET AL, "RNA-GUIDED HUMAN GENOME ENGINEERING VIA CAS9", SCIENCE, (20130215), vol. 339, no. 6121, doi:10.1126/SCIENCE.1232033, pages 823 - 826, XP055111247, DOI:   http://dx.doi.org/10.1126/science.1232033

D4:WOONG Y HWANG ET AL, "EFFICIENT GENOME EDITING IN ZEBRAFISH USING A CRISPR-CAS SYSTEM", NATURE BIOTECHNOLOGY, (201303), vol. 31, no. 3, doi:10.1038/NBT.2501, pages 227 - 229, XP055086625, DOI:   http://dx.doi.org/10.1038/nbt.2501

 

EP2784162:特許取消(2019/02/13)→審判請求(2019/02/15)

1.  An engineered, non-naturally occurring Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR)-CRISPR associated (Cas) (CRISPR-Cas) vector system comprising one or more vectors comprising:

  a) a first regulatory element operably linked to one or more nucleotide sequences encoding one or more CRISPR-Cas system polynucleotide sequences comprising a guide sequence, a tracr RNA, and a tracr mate sequence, wherein the guide sequence hybridizes with one or more target sequences in polynucleotide loci in a eukaryotic cell ,

  b) a second regulatory element operably linked to a nucleotide sequence encoding a Type II Cas9 protein, wherein components (a) and (b) are located on same or different vectors of the system,

  wherein the CRISPR-Cas system comprises two or more nuclear localization signals (NLSs) expressed with the nucleotide sequence encoding the Cas9 protein,

   whereby the one or more guide sequences target the one or more polynucleotide loci in a eukaryotic cell and the Cas9 protein cleaves the one or more polynucleotide loci,

   whereby the sequence of the one or more polynucleotide loci is modified.

 

・こちらも優先権が争われていたので、優先権無効→新規性or進歩性欠如か?

 

EP2825654:口頭審理未定(補正クレーム提出済(2018/09/03))

1.  A non-naturally occurring or engineered composition comprising a vector system comprising one or more vectors comprising

  I. a first regulatory element operably linked to a CRISPR-Cas system chimeric RNA (chiRNA) polynucleotide sequence, wherein the polynucleotide sequence comprises

    (a) a guide sequence capable of hybridizing to a target sequence in a eukaryotic cell,     (b) a tracr mate sequence, and

    (c) a tracr sequence, and

  II. a second regulatory element operably linked to an enzyme-coding sequence encoding a CRISPR enzyme comprising at least one or more nuclear localization sequences (NLSs) in the proximity of a terminus of the CRISPR enzyme, wherein (a), (b) and (c) are arranged in a 5' to 3' orientation,

  wherein components I and II are located on the same or different vectors of the system,

  wherein when transcribed, the tracr mate sequence hybridizes to the tracr sequence and the guide sequence directs sequence-specific binding of a CRISPR complex to the target sequence,

   wherein the CRISPR complex comprises the CRISPR enzyme complexed with (1) the guide sequence that is hybridized to the target sequence, and (2) the tracr mate sequence that is hybridized to the tracr sequence, and

   wherein the chimeric RNA polynucleotide sequence comprises two or more hairpins.

 

EP2840140:口頭審理未定(補正クレーム提出済(2018/07/23))

1.  A method of selecting one or more prokaryotic cell(s) by introducing one or more mutations in one or more prokaryotic cell(s), the method comprising:

   introducing one or more vectors into the prokaryotic cell(s);

   wherein the one or more vectors drive expression of one or more of: a CRISPR enzyme, a guide sequence linked to a tracr mate sequence, a tracr sequence, and an editing template for recombination into a targeted chromosomal locus comprising a target polynucleotide;   and all of a CRISPR enzyme, a guide sequence linked to a tracr mate sequence, and a tracr sequence, and an editing template are produced in the prokaryotic cell(s);

   introducing the editing template into a target polynucleotide through recombination,

   wherein the editing template comprises the one or more mutations of the target polynucleotide that alter either the protospacer-adjacent motif (PAM) sequence or the protospacer sequence, and that abolish CRISPR enzyme cleavage of the target polynucleotide;

   allowing a CRISPR complex to bind to the target polynucleotide to effect cleavage of the target polynucleotide;

   wherein the CRISPR complex comprises the CRISPR enzyme complexed with (1) the guide sequence that is hybridized to a target sequence within the target polynucleotide, and (2) the tracr mate sequence that is hybridized to a tracr sequence;

   wherein cleavage of the target polynucleotide by the CRISPR complex induces cell death;

   thereby allowing one or more prokaryotic cell(s) in which one or more mutations have been introduced to be selected.

 

・対象が原核生物(重要性低)

 

EP2931892:特許許可(2018/08/06)

1.  A computer implemented method for selecting a CRISPR complex for targeting and/or cleavage of a candidate target nucleic acid sequence within a cell, comprising the steps of:

  (a) determining amount, location and nature of mismatch(es) of guide sequence of potential CRISPR complex(es) and the candidate target nucleic acid sequence,

  (b) determining contribution of each of the amount, location and nature of mismatch(es) to hybridization free energy of binding between the target nucleic acid sequence and the guide sequence of potential CRISPR complex(es) from a training data set,

  (c) based on the contribution analysis of step (b), predicting cleavage at the location(s) of the mismatch(es) of the target nucleic acid sequence by the potential CRISPR complex(es), and

  (d) selecting the CRISPR complex from potential CRISPR complex(es) based on whether the prediction of step (c) indicates that it is more likely than not that cleavage will occur at location(s) of mismatch(es) by the CRISPR complex wherein step (b) is performed by defining a thermodynamic model having a set of weights linking effective free energy of hybridization Z to local free energies G;

   defining a training set of guide RNA/target DNA sequence pairs; inputting known values of local free energies G for each guide RNA/target DNA sequence pair in the training set; calculating a value of effective free energy of hybridization Z for each guide RNA/target DNA sequence pair in the training set;

   determining the weights using a machine learning algorithm and outputting the weights whereby the weights can be used to estimate the free energy of hybridization for any sequence.

 

機械学習を用いた設計用プログラム

 

EP2931897:口頭審理未定(補正クレーム提出済(2019/01/08))

1.  A composition comprising a CRISPR complex comprising

  a) a tracr mate RNA polynucleotide,

  b) a tracr RNA polynucleotide,

  c) a guide RNA polynucleotide capable of hybridizing to a target sequence in a eukaryotic cell or a cell in a multi-cellular eukaryotic organism; and

  d) a Cas9 comprising one or more nuclear localization sequences, for use in therapy.

 

・CRISPR-Cas9複合体を含む第一医薬用途組成物。なぜこのクレームで通る…。

 

EP2898075:口頭審理待ち(2019/07/04、補正クレーム提出済(2018/05/09))

1.  A composition comprising:

  CRISPR complex components comprising:

       I. CRISPR-Cas system polynucleotide sequence(s) which comprise(s):

           (a) an engineered guide sequence comprised of RNA and capable of hybridizing to a target sequence in a polynucleotide locus,

           (b) a tracr mate sequence comprised of RNA, and

           (c) a tracrRNA sequence comprised of RNA, and wherein (a), (b) and (c) are arranged in a 5' to 3' orientation,

       II. a Type II Cas9 protein, wherein the tracr mate sequence hybridizes to the tracrRNA sequence and the guide sequence directs sequence-specific binding of a CRISPR complex to a target sequence,

  wherein the CRISPR complex comprises the Type II Cas9 protein complexed with (1) the guide sequence that is hybridized to the target sequence, and (2) the tracr mate sequence that is hybridized to the tracrRNA sequence, and

  wherein the Type II Cas9 protein is or comprises a Staphylococcus aureus Cas9 (SaCas9).

 

・SaCas9タイプ(小さいCas9)のII型CRISPR-Cas9複合体、JPに対応する特許があったはず。

 

EP2931898:口頭審理待ち(2019/05/15、補正クレーム提出済(2018/03/27))

1.  A non-naturally occurring or engineered composition comprising:

  a delivery system operably configured to deliver CRISPR-Cas complex components or polynucleotide sequences comprising or encoding said components into a eukaryotic cell, wherein said CRISPR-Cas complex is operable in the eukaryotic cell;

       I. one or more CRISPR-Cas complex polynucleotide sequences comprising or encoding for expression in the eukaryotic cell:

           (a) a guide sequence capable of hybridizing to a target sequence in a eukaryotic cell,

           (b) a tracr mate sequence, and

           (c) a tracr sequence, and

       II. a CRISPR enzyme comprising one or more heterologous functional domains or a polynucleotide encoding a CRISPR enzyme comprising one or more heterologous functional domains for expression in the eukaryotic cell; wherein:

    the tracr mate sequence hybridizes to the tracr sequence,

    the guide sequence directs sequence-specific binding of a CRISPR complex to the target sequence,

    the CRISPR complex comprises the CRISPR enzyme complexed with (1) the guide sequence that is hybridized to the target sequence, and (2) the tracr mate sequence that is hybridized to the tracr sequence,

    the CRISPR enzyme comprises one or more mutations, such that the enzyme has altered nuclease activity compared with the wild type enzyme,

    wherein the one or more heterologous functional domains comprises at least one or more Nuclear Localization Signals NLS(s).

 

・Cas9以外のCas(CRISPR enzyme)を包含する特許がついに…。サポート要件との兼ね合いがどうなるか。UC Berkeleyの出願とは異なり、パイオニア発明では無いので、もっと厳しく判断されてもいいものであるが。