JPWO2019217944A5 - - Google Patents
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- JPWO2019217944A5 JPWO2019217944A5 JP2021513765A JP2021513765A JPWO2019217944A5 JP WO2019217944 A5 JPWO2019217944 A5 JP WO2019217944A5 JP 2021513765 A JP2021513765 A JP 2021513765A JP 2021513765 A JP2021513765 A JP 2021513765A JP WO2019217944 A5 JPWO2019217944 A5 JP WO2019217944A5
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- rtt
- mecp2
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- 101700029603 MECP2 Proteins 0.000 claims 34
- 235000001014 amino acid Nutrition 0.000 claims 25
- 102100014726 MECP2 Human genes 0.000 claims 24
- 229920000023 polynucleotide Polymers 0.000 claims 23
- 239000002157 polynucleotide Substances 0.000 claims 23
- 125000003275 alpha amino acid group Chemical group 0.000 claims 21
- 238000006467 substitution reaction Methods 0.000 claims 21
- 101700056051 cas9 Proteins 0.000 claims 18
- 229920001850 Nucleic acid sequence Polymers 0.000 claims 15
- 150000007523 nucleic acids Chemical group 0.000 claims 15
- 210000004027 cells Anatomy 0.000 claims 13
- 230000004568 DNA-binding Effects 0.000 claims 12
- 230000000875 corresponding Effects 0.000 claims 11
- 102000009914 Adenosine deaminases Human genes 0.000 claims 9
- 108091022188 Adenosine deaminases Proteins 0.000 claims 9
- 239000004475 Arginine Substances 0.000 claims 9
- 229920005681 CRISPR RNA Polymers 0.000 claims 9
- 230000004048 modification Effects 0.000 claims 9
- 238000006011 modification reaction Methods 0.000 claims 9
- 229920001184 polypeptide Polymers 0.000 claims 7
- 229920002391 Guide RNA Polymers 0.000 claims 6
- 108020005004 Guide RNA Proteins 0.000 claims 6
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims 6
- 239000004473 Threonine Substances 0.000 claims 6
- 230000000295 complement Effects 0.000 claims 6
- 229920003013 deoxyribonucleic acid Polymers 0.000 claims 6
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 claims 5
- 150000001413 amino acids Chemical class 0.000 claims 4
- OIRDTQYFTABQOQ-GAWUUDPSSA-N 9-β-D-XYLOFURANOSYL-ADENINE Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@H](O)[C@H]1O OIRDTQYFTABQOQ-GAWUUDPSSA-N 0.000 claims 3
- OIRDTQYFTABQOQ-SXVXDFOESA-N Adenosine Natural products Nc1ncnc2c1ncn2[C@@H]3O[C@@H](CO)[C@H](O)[C@@H]3O OIRDTQYFTABQOQ-SXVXDFOESA-N 0.000 claims 3
- 229920000195 Bacterial small RNA Polymers 0.000 claims 3
- 239000002126 C01EB10 - Adenosine Substances 0.000 claims 3
- 229920000033 CRISPR Polymers 0.000 claims 3
- 108010082319 CRISPR-Associated Protein 9 Proteins 0.000 claims 3
- 102200028136 CSN1S1 D10A Human genes 0.000 claims 3
- 108020004705 Codon Proteins 0.000 claims 3
- XUJNEKJLAYXESH-REOHCLBHSA-N L-cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims 3
- 108010072388 Methyl-CpG-Binding Protein 2 Proteins 0.000 claims 3
- 241000193996 Streptococcus pyogenes Species 0.000 claims 3
- 229940076156 Streptococcus pyogenes Drugs 0.000 claims 3
- 229960005305 adenosine Drugs 0.000 claims 3
- 235000018417 cysteine Nutrition 0.000 claims 3
- 210000002569 neurons Anatomy 0.000 claims 3
- 239000002773 nucleotide Substances 0.000 claims 2
- 125000003729 nucleotide group Chemical group 0.000 claims 2
- 241000124008 Mammalia Species 0.000 claims 1
- 238000000338 in vitro Methods 0.000 claims 1
- 210000004962 mammalian cells Anatomy 0.000 claims 1
- OZAIFHULBGXAKX-UHFFFAOYSA-N precursor Substances N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 claims 1
- 201000010874 syndrome Diseases 0.000 claims 1
- 125000000341 threoninyl group Chemical group [H]OC([H])(C([H])([H])[H])C([H])(N([H])[H])C(*)=O 0.000 claims 1
Claims (15)
ｉｉ）RTTに関連するSNPにおける前記A・TからG・Cへの改変が、メチルCpG結合タンパク質2（Mecp2）ポリペプチドにおいてシステインをアルギニンに、メチオニンをスレオニンに、または停止コドンをアルギニンに変更し、および／または
ｉｉｉ）RTTに関連する前記SNPが、アミノ酸168位、133位、255位、270位、もしくは306位にアルギニン、または158位にスレオニンを含むMecp2ポリペプチドの発現をもたらし、および／または
ｉｖ）前記ポリヌクレオチドプログラム可能なDNA結合ドメインが、Streptococcus pyogenesのCas9（SpCas9）またはそのバリアントであり、および／または
ｖ）前記ポリヌクレオチドプログラム可能なDNA結合ドメインが、改変されたプロトスペーサー隣接モチーフ（PAM）特異性を有する改変されたSpCas9を含み、好ましくは
ｖ－１）前記改変されたSpCas9が、核酸配列5'-NGT-3'に対する特異性を有し、より好ましくは
ｖ－２）前記改変されたSpCas9が、アミノ酸置換L1111R、D1135V、G1218R、E1219F、A1322R、R1335V、T1337R、ならびにL1111、D1135L、S1136R、G1218S、E1219V、D1332A、D1332S、D1332T、D1332V、D1332L、D1332K、D1332R、R1335Q、T1337、T1337L、T1337Q、T1337I、T1337V、T1337F、T1337S、T1337N、T1337K、T1337H、T1337Q、およびT1337Mのうち1つもしくは複数、またはその対応するアミノ酸置換を含むか、または
前記改変されたSpCas9が、アミノ酸置換D1135L、S1136R、G1218S、E1219V、A1322R、R1335Q、およびT1337、ならびにL1111R、G1218R、E1219F、D1332A、D1332S、D1332T、D1332V、D1332L、D1332K、D1332R、T1337L、T1337I、T1337V、T1337F、T1337S、T1337N、T1337K、T1337R、T1337H、T1337Q、およびT1337Mのうち1つもしくは複数、またはその対応するアミノ酸置換を含み、および／または
ｖｉ）前記ポリヌクレオチドプログラム可能なDNA結合ドメインが、ヌクレアーゼ不活性バリアントまたはニッカーゼバリアントであり、好ましくは前記ニッカーゼバリアントが、アミノ酸置換D10Aまたはその対応するアミノ酸置換を含み、および／または
ｖｉｉ）
ｖｉｉ－１）前記アデノシンデアミナーゼドメインが、デオキシリボ核酸（DNA）中のアデノシンを脱アミノ化することができ、好ましくは
ｖｉｉ－２）前記アデノシンデアミナーゼが、TadAデアミナーゼであり、および／または前記TadAデアミナーゼが、TadA*7.10であり、および／または
ｖｉｉｉ）前記1つまたは複数のガイドRNAが、CRISPR RNA（crRNA）およびトランスコード化小RNA（tracrRNA）を含み、前記crRNAが、RTTに関連するSNPを含むMecp2核酸配列に相補的な核酸配列を含み、および／または
前記塩基エディターが、RTTに関連するSNPを含むMecp2核酸配列に相補的な核酸配列を含むシングルガイドRNA（sgRNA）と複合体化している、
請求項１に記載の方法。 i) The modification is one or more of R106W, R168 *, R133C, T158M, R255 *, R270 *, and R306C , and / or
ii) The above-mentioned A / T to G / C modification in RTT-related SNPs changed cysteine to arginine, methionine to threonine, or stop codon to arginine in the methylCpG-binding protein 2 (Mecp2) polypeptide. , And / or
iii) The SNP associated with RTT results in the expression of a Mecp2 polypeptide containing arginine at amino acids 168, 133, 255, 270, or 306, or threonine at 158 , and / or.
iv) The polynucleotide programmable DNA-binding domain is Cas9 (SpCas9) or a variant thereof of Streptococcus pyogenes and / or
v) The polynucleotide programmable DNA binding domain comprises modified SpCas9 with modified protospacer flanking motif (PAM) specificity , preferably.
v-1) The modified SpCas9 has specificity for the nucleic acid sequence 5'-NGT-3' , and is more preferable.
v-2) The modified SpCas9 contains amino acid substitutions L1111R, D1135V, G1218R, E1219F, A1322R, R1335V, T1337R, and L1111, D1135L, S1136R, G1218S, E1219V, D1332A, D1332S, D1332T, D1332V, Contains or contains one or more of D1332R, R1335Q, T1337, T1337L, T1337Q, T1337I, T1337V, T1337F, T1337S, T1337N, T1337K, T1337H, T1337Q, and T1337M, or their corresponding amino acid substitutions.
The modified SpCas9 contains amino acid substitutions D1135L, S1136R, G1218S, E1219V, A1322R, R1335Q, and T1337, as well as L1111R, G1218R, E1219F, D1332A, D1332S, D1332T, D1332V, D1332L, D1332K, D1332T. , T1337F, T1337S, T1337N, T1337K, T1337R, T1337H, T1337Q, and T1337M containing one or more of, or their corresponding amino acid substitutions , and / or
vi) The polynucleotide programmable DNA binding domain is a nuclease-inactivated variant or a nickase variant , preferably the nickase variant comprises amino acid substitution D10A or its corresponding amino acid substitution and / or.
vii)
vii-1) The adenosine deaminase domain can deaminate adenosine in deoxyribonucleic acid (DNA) , preferably.
vii-2) The adenosine deaminase is TadA deaminase and / or the TadA deaminase is TadA * 7.10 and / or
viii) The one or more guide RNAs contain a CRISPR RNA (crRNA) and a transcoded small RNA (tracrRNA), the crRNA having a nucleic acid sequence complementary to the Mecp2 nucleic acid sequence containing the SNP associated with RTT. Including and / or
The base editor is complexed with a single guide RNA (sgRNA) containing a nucleic acid sequence complementary to the Mecp2 nucleic acid sequence containing the SNP associated with RTT.
The method according to claim 1 .
前記塩基エディターをターゲティングしてRTTに関連するSNPのA・TからG・Cへの改変をもたらす1つもしくは複数のガイドポリヌクレオチド
を含む、細胞またはその前駆体。 Polynucleotides A / T-to-G / C modification of RTT-related SNPs by targeting a base editor containing a programmable DNA-binding domain and an adenosindeaminase domain or a polynucleotide encoding the base editor, as well as the base editor. A cell or precursor thereof, comprising one or more guide polynucleotides that result in .
ｉｉ）前記ニューロンはMecp2ポリペプチドを発現する、
請求項３に記載の細胞。 i) The cell is a neuron and / or
ii) The neuron expresses a Mecp2 polypeptide,
The cell according to claim 3 .
ｉｉ）前記細胞は哺乳動物細胞であり、および／または
ｉｉｉ）前記細胞はヒト細胞であり、および／または
ｉｖ）前記改変が、R106W、R168*、R133C、T158M、R255*、R270*、およびR306Cのうち1つまたは複数であり、および／または
ｖ）RTTに関連するSNPにおける前記A・TからG・Cへの改変が、メチルCpG結合タンパク質2（Mecp2）ポリペプチドにおいてシステインをアルギニンに、メチオニンをスレオニンに、または停止コドンをアルギニンに変更し、および／または
ｖｉ）RTTに関連するSNPが、アミノ酸168位、133位、255位、270位、もしくは306位にアルギニン、または158位にスレオニンを含むMecp2ポリペプチドの発現をもたらし、および／または
ｖｉｉ）前記ポリヌクレオチドプログラム可能なDNA結合ドメインが、Streptococcus pyogenesのCas9（SpCas9）またはそのバリアントであり、および／または
ｖｉｉｉ）前記ポリヌクレオチドプログラム可能なDNA結合ドメインが、改変されたプロトスペーサー隣接モチーフ（PAM）特異性を有する改変されたSpCas9を含み、好ましくは前記改変されたSpCas9が、核酸配列5'-NGT-3'に対する特異性を有し、より好ましくは前記改変されたSpCas9が、アミノ酸置換L1111R、D1135V、G1218R、E1219F、A1322R、R1335V、T1337R、ならびにL1111、D1135L、S1136R、G1218S、E1219V、D1332A、D1332S、D1332T、D1332V、D1332L、D1332K、D1332R、R1335Q、T1337、T1337L、T1337Q、T1337I、T1337V、T1337F、T1337S、T1337N、T1337K、T1337H、T1337Q、およびT1337Mのうち1つもしくは複数、またはその対応するアミノ酸置換を含むか、または前記改変されたSpCas9が、アミノ酸置換D1135L、S1136R、G1218S、E1219V、A1322R、R1335Q、およびT1337、ならびにL1111R、G1218R、E1219F、D1332A、D1332S、D1332T、D1332V、D1332L、D1332K、D1332R、T1337L、T1337I、T1337V、T1337F、T1337S、T1337N、T1337K、T1337R、T1337H、T1337Q、およびT1337Mのうち1つもしくは複数、またはその対応するアミノ酸置換を含む、
請求項３または４に記載の細胞。 i) The cells are from a subject with RTT and / or
ii) The cells are mammalian cells and / or
iii) The cells are human cells and / or
iv) The modification is one or more of R106W, R168 *, R133C, T158M, R255 *, R270 *, and R306C , and / or
v) The above-mentioned A / T to G / C modification in RTT-related SNPs changed cysteine to arginine, methionine to threonine, or stop codon to arginine in the methyl CpG-binding protein 2 (Mecp2) polypeptide. , And / or
vi) The SNP associated with RTT results in the expression of a Mecp2 polypeptide containing arginine at amino acids 168, 133, 255, 270, or 306, or threonine at 158 , and / or.
vii) The polynucleotide programmable DNA-binding domain is Cas9 (SpCas9) of Streptococcus pyogenes or a variant thereof , and / or.
viii) The polynucleotide programmable DNA binding domain comprises modified SpCas9 with modified protospacer flanking motif (PAM) specificity , preferably said modified SpCas9 is nucleic acid sequence 5'-NGT. The modified SpCas9, which has specificity for -3'and more preferably the above-mentioned modified SpCas9, has amino acid substitutions L1111R, D1135V, G1218R, E1219F, A1322R, R1335V, T1337R, and L1111, D1135L, S1136R, G1218S, E1219V, D1332A, D1332S. , D1332T, D1332V, D1332L, D1332K, D1332R, R1335Q, T1337, T1337L, T1337Q, T1337I, T1337V, T1337F, T1337S, T1337N, T1337K, T1337H, T1337Q, and one or more of the corresponding amino acids, T1337Q, and T1337M. , Or the modified SpCas9 described above, contains the amino acid substitutions D1135L, S1136R, G1218S, E1219V, A1322R, R1335Q, and T1337, as well as L1111R, G1218R, E1219F, D1332A, D1332S, D1332T, D1332V, D1332L, D1332K. Includes one or more of T1337L, T1337I, T1337V, T1337F, T1337S, T1337N, T1337K, T1337R, T1337H, T1337Q, and T1337M, or their corresponding amino acid substitutions.
The cell according to claim 3 or 4 .
ｉｉ）前記アデノシンデアミナーゼドメインが、デオキシリボ核酸（DNA）中のアデノシンを脱アミノ化することができ、好ましくは
ｉｉ－１）前記アデノシンデアミナーゼが、TadAデアミナーゼであり、および／または
ｉｉ－２）前記TadAデアミナーゼが、TadA*7.10であり、および／または
ｉｉｉ）前記1つまたは複数のガイドRNAが、CRISPR RNA（crRNA）およびトランスコード化小RNA（tracrRNA）を含み、前記crRNAが、RTTに関連するSNPを含むMecp2核酸配列に相補的な核酸配列を含み、および／または
ｉｖ）前記塩基エディターが、RTTに関連するSNPを含むMecp2核酸配列に相補的な核酸配列を含むシングルガイドRNA（sgRNA）と複合体化している、
請求項３～５のいずれか一項に記載の細胞。 i) The polynucleotide programmable DNA binding domain is a nuclease-inert variant or nickase variant , preferably said nickase variant comprising amino acid substitution D10A or its corresponding amino acid substitution and / or.
ii) The adenosine deaminase domain can deaminate adenosine in deoxyribonucleic acid (DNA) , preferably.
ii-1) The adenosine deaminase is TadA deaminase and / or
ii-2) The TadA deaminase is TadA * 7.10 and / or
iii) The one or more guide RNAs contain a CRISPR RNA (crRNA) and a transcoded small RNA (tracrRNA), the crRNA having a nucleic acid sequence complementary to the Mecp2 nucleic acid sequence containing the SNP associated with RTT. Including and / or
iv) The base editor is complexed with a single guide RNA (sgRNA) containing a nucleic acid sequence complementary to a Mecp2 nucleic acid sequence containing an SNP associated with RTT.
The cell according to any one of claims 3 to 5 .
ポリヌクレオチドプログラム可能なDNA結合ドメインおよびアデノシンデアミナーゼドメインを含む前記塩基エディターまたは前記塩基エディターをコードするポリヌクレオチド、および
前記塩基エディターをターゲティングしてRTTに関連するSNPのA・TからG・Cへの改変をもたらす1つもしくは複数のガイドポリヌクレオチド
を前記対象に投与することを含む、
組成物。 A composition comprising a base editor or a polynucleotide encoding it for use in a method of treating RTT in a subject, said method.
Polynucleotides The nucleotides that contain programmable DNA-binding domains and adenosine deaminase domains, or polynucleotides that encode the base editors, and targeting the base editors for RTT-related SNPs from A / T to G / C. Containing administration of one or more guide polynucleotides that result in modification to said subject.
Composition .
ｉｉ）前記方法は、前記塩基エディターまたはそれをコードするポリヌクレオチドと、前記1つもしくは複数のガイドポリヌクレオチドとを、前記対象の細胞に送達することを含む、
請求項８に記載の組成物。 i) The subject is a mammal or human and / or
ii) The method comprises delivering the base editor or a polynucleotide encoding it and the one or more guide polynucleotides to the cell of interest.
The composition according to claim 8 .
ｉｉ）前記改変が、R106W、R168*、R133C、T158M、R255*、R270*、およびR306Cのうち1つまたは複数であり、および／または
ｉｉｉ）RTTに関連するSNPにおける前記A・TからG・Cへの改変が、メチルCpG結合タンパク質2（Mecp2）ポリペプチドにおいてシステインをアルギニンに、メチオニンをスレオニンに、または停止コドンをアルギニンに変更し、および／または
ｉｖ）RTTに関連するSNPが、アミノ酸168位、133位、255位、270位、もしくは306位にアルギニン、または158位にスレオニンを含むMecp2ポリペプチドの発現をもたらし、および／または
ｖ）前記ポリヌクレオチドプログラム可能なDNA結合ドメインが、Streptococcus pyogenesのCas9（SpCas9）またはそのバリアントであり、および／または
ｖｉ）前記ポリヌクレオチドプログラム可能なDNA結合ドメインが、改変されたプロトスペーサー隣接モチーフ（PAM）特異性を有する改変されたSpCas9を含み、好ましくは前記改変されたSpCas9が、核酸配列5'-NGT-3'に対する特異性を有し、より好ましくは
前記改変されたSpCas9が、アミノ酸置換L1111R、D1135V、G1218R、E1219F、A1322R、R1335V、T1337R、ならびにL1111、D1135L、S1136R、G1218S、E1219V、D1332A、R1335Q、T1337、T1337L、T1337Q、T1337I、T1337V、T1337F、およびT1337Mのうち1つもしくは複数、またはその対応するアミノ酸置換を含むか、もしくは
前記改変されたSpCas9が、アミノ酸置換D1135L、S1136R、G1218S、E1219V、A1322R、R1335Q、およびT1337、ならびにL1111R、D1135L、S1136R、G1218S、E1219V、D1332A、D1332S、D1332T、D1332V、D1332L、D1332K、D1332R、R1335Q、T1337、T1337L、T1337Q、T1337I、T1337V、T1337F、T1337S、T1337N、T1337K、T1337R、T1337H、T1337Q、およびT1337Mのうち1つもしくは複数、またはその対応するアミノ酸置換を含む、
請求項８または９に記載の組成物。 i) The cell is a neuron and / or
ii) The modification is one or more of R106W, R168 *, R133C, T158M, R255 *, R270 *, and R306C , and / or
iii) The above-mentioned A / T to G / C modification in RTT-related SNPs changed cysteine to arginine, methionine to threonine, or stop codon to arginine in the methylCpG-binding protein 2 (Mecp2) polypeptide. , And / or
iv) RTT-related SNPs result in the expression of a Mecp2 polypeptide containing arginine at amino acids 168, 133, 255, 270, or 306, or threonine at position 158 , and / or.
v) The polynucleotide programmable DNA binding domain is Cas9 (SpCas9) or a variant thereof of Streptococcus pyogenes and / or
vi) The polynucleotide programmable DNA binding domain comprises modified SpCas9 with modified protospacer flanking motif (PAM) specificity , preferably said modified SpCas9 is nucleic acid sequence 5'-NGT. Has specificity for -3' , more preferably
The modified SpCas9 contains amino acid substitutions L1111R, D1135V, G1218R, E1219F, A1322R, R1335V, T1337R, and L1111, D1135L, S1136R, G1218S, E1219V, D1332A, R1335Q, T1337, T1337L, T1337Q, T1337L, T1337T. And one or more of T1337M, or their corresponding amino acid substitutions , or
The modified SpCas9 contains amino acid substitutions D1135L, S1136R, G1218S, E1219V, A1322R, R1335Q, and T1337, as well as L1111R, D1135L, S1136R, G1218S, E1219V, D1332A, D1332S, D1332T, D1332V, D1332L. , T1337, T1337L, T1337Q, T1337I, T1337V, T1337F, T1337S, T1337N, T1337K, T1337R, T1337H, T1337Q, and T1337M, including one or more, or their corresponding amino acid substitutions.
The composition according to claim 8 or 9 .
ｉ－１）前記ニッカーゼバリアントが、アミノ酸置換D10Aまたはその対応するアミノ酸置換を含み、および／または
ｉｉ）前記アデノシンデアミナーゼドメインが、デオキシリボ核酸（DNA）中のアデノシンを脱アミノ化することができる、
請求項８～１０のいずれか一項に記載の組成物。 i) The polynucleotide programmable DNA binding domain is a nuclease-inactivated variant or a nickase variant , preferably
i-1) The nickase variant comprises an amino acid substitution D10A or its corresponding amino acid substitution and / or
ii) The adenosine deaminase domain can deaminate adenosine in deoxyribonucleic acid (DNA),
The composition according to any one of claims 8 to 10 .
ｉｉ）前記1つまたは複数のガイドRNAが、CRISPR RNA（crRNA）およびトランスコード化小RNA（tracrRNA）を含み、前記crRNAが、RTTに関連するSNPを含むMecp2核酸配列に相補的な核酸配列を含み、および／または
ｉｉｉ）前記塩基エディターが、RTTに関連するSNPを含むMecp2核酸配列に相補的な核酸配列を含むシングルガイドRNA（sgRNA）と複合体化している、
請求項１１または１２に記載の組成物。 i) The TadA deaminase is TadA * 7.10 and / or
ii) The one or more guide RNAs contain a CRISPR RNA (crRNA) and a transcoded small RNA (tracrRNA), the crRNA having a nucleic acid sequence complementary to the Mecp2 nucleic acid sequence containing the SNP associated with RTT. Including and / or
iii) The base editor is complexed with a single guide RNA (sgRNA) containing a nucleic acid sequence complementary to a Mecp2 nucleic acid sequence containing an SNP associated with RTT.
The composition according to claim 11 or 12 .
(ii)アデノシンデアミナーゼと
を含む、塩基エディター。 (i) Amino acid substitutions L1111R, D1135V, G1218R, E1219F, A1322R, R1335V, T1337R, and L1111, D1135L, S1136R, G1218S, E1219V, D1332A, D1332S, D1332T, D1332V, D1332L, D1332T Modified SpCas9 containing one or more of T1337Q, T1337I, T1337V, T1337F, T1337S, T1337N, T1337K, T1337H, T1337Q, and T1337M, or their corresponding amino acid substitutions, and
(ii) A base editor containing adenosine deaminase.
(ii)アデノシンデアミナーゼと
を含む、塩基エディター。
(i) Amino acid substitutions D1135L, S1136R, G1218S, E1219V, A1322R, R1335Q, and T1337, as well as L1111R, G1218R, E1219F, D1332A, D1332S, D1332T, D1332V, D1332L, D1332K, D1332R, T1337L, D1332R, T1337L , T1337N, T1337K, T1337R, T1337H, T1337Q, and T1337M with one or more of the modified SpCas9, including one or more of them, or their corresponding amino acid substitutions.
(ii) A base editor containing adenosine deaminase.
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