IP Library › Patent Application 17641343
Patent Application
App. No. 17/641,343

NOVEL NUCLEOBASE EDITORS AND METHODS OF USING SAME

Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US None
App. No.
17/641,343
Abstract

The invention features novel programmable nucleobase editors comprising adenosine deaminase domains and methods of using the same for polynucleotide editing. In some embodiments, programmable nucleobase editors edit a pathogenic mutation associated with a genetic disease.

Claims (299)

1 . An adenosine deaminase comprising an alteration at an amino acid position selected from the group consisting of 21, 23, 25, 38, 51, 54, 70, 71, 72, 73, 94, 124, 133, 139, 146, and 158 of SEQ ID NO: 1, or a corresponding alteration in another adenosine deaminase:

(SEQ ID NO: 1)

        10         20         30         40

MSEVEFSHEY WMRHALTLAK  R A R D E REVPV GAVLVLN N RV

        50         60         70         80

IGEGWNRAIG  L HD P TAHAEI MALRQGGLV M   QNY RLIDATL

        90        100        110        120

YVTFEPCVMC AGA M IHSRIG RVVFGVRNAK TGAAGSLMDV

       130        140        150        160

LHYPGMNHRV EI T EGILA D E GAALL C YFER MPRQVFN A QK

KAQSSTD.

2 . The adenosine deaminase of claim 1 , which comprises an alteration selected from the group consisting of R21N, R23H, E25F, N38G, L51W, P54C, M70V, Q71M, N72K, Y73S, M94V, P124W, T133K, D139L, D139M, C146R, and A158K of SEQ ID NO: 1, or a corresponding alteration in another adenosine deaminase.

3 . The adenosine deaminase of claim 1 , further comprising a V82T alteration of SEQ ID NO: 1, or a corresponding alteration in another adenosine deaminase.

4 - 6 . (canceled)

7 . The adenosine deaminase of claim 1 , which further comprises one or more of the following alterations: Y147T, Y147R, Q154S, Y123H, and Q154R.

8 . The adenosine deaminase of claim 1 , wherein the adenosine deaminase comprises any one of the following groups of alterations:

E25F+V82S+Y123H;

T133K+Y147R+Q154R;

E25F+V82S+Y123H+Y147R+Q154R;

L51W+V82S+Y123H+C146R+Y147R+Q154R;

Y73S+V82S+Y123H+Y147R+Q154R;

P54C+V82S+Y123H+Y147R+Q154R;

N38G+V82T+Y123H+Y147R+Q154R;

N72K+V82S+Y123H+D139L+Y147R+Q154R;

E25F+V82S+Y123H+D139M+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

E25F+V82S+Y123H+T133K+Y147R+Q154R;

E25F+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+P124W+Y147R+Q154R;

L51W+V82S+Y123H+C146R+Y147R+Q154R;

P54C+V82S+Y123H+Y147R+Q154R;

Y73S+V82S+Y123H+Y147R+Q154R;

N38G+V82T+Y123H+Y147R+Q154R;

R23H+V82S+Y123H+Y147R+Q154R;

R21N+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+Y147R+Q154R+A158K;

N72K+V82S+Y123H+D139L+Y147R+Q154R;

E25F+V82S+Y123H+D139M+Y147R+Q154R;

M70V+V82S+M94V+Y123H+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

E25F+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82T+Y123H+Y147R+Q154R;

N38G+I76Y+V82S+Y123H+Y147R+Q154R;

R23H+I76Y+V82S+Y123H+Y147R+Q154R;

P54C+I76Y+V82S+Y123H+Y147R+Q154R;

R21N+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82S+Y123H+D139M+Y147R+Q154R;

Y73S+I76Y+V82S+Y123H+Y147R+Q154R;

E25F+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82T+Y123H+Y147R+Q154R;

N38G+I76Y+V82S+Y123H+Y147R+Q154R;

R23H+I76Y+V82S+Y123H+Y147R+Q154R;

P54C+I76Y+V82S+Y123H+Y147R+Q154R;

R21N+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82S+Y123H+D139M+Y147R+Q154R;

Y73S+I76Y+V82S+Y123H+Y147R+Q154R;

V82S+Q154R;

N72K+V82S+Y123H+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R+A158K;

M70V+Q71M+N72K+V82S+Y123H+Y147R+Q154R;

N72K V82S+Y123H+Y147R+Q154R;

Q71M V82S+Y123H+Y147R+Q154R;

M70V+V82S+M94V+Y123H+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R+A158K; or

M70V+Q71M+N72K+V82S+Y123H+Y147R+Q154R.

9 - 11 . (canceled)

12 . A fusion protein comprising a polynucleotide programmable DNA binding domain and at least one base editor domain that is an adenosine deaminase variant comprising an alteration at an amino acid position selected from the group consisting of 21, 23, 25, 38, 51, 54, 70, 71, 72, 73, 94, 124, 133, 139, 146, and 158 of SEQ ID NO: 1, or a corresponding alteration in another adenosine deaminase:

(SEQ ID NO: 1)

        10         20         30         40

MSEVEFSHEY WMRHALTLAK  R A R D E REVPV GAVLVLN N RV

        50         60         70         80

IGEGWNRAIG  L HD P TAHAEI MALRQGGLV M   QNY RLIDATL

        90        100        110        120

YVTFEPCVMC AGA M IHSRIG RVVFGVRNAK TGAAGSLMDV

       130        140        150        160

LHYPGMNHRV EI T EGILA D E GAALL C YFER MPRQVFN A QK

KAQSSTD.

13 . (canceled)

14 . A fusion protein comprising a polynucleotide programmable DNA binding domain and at least one base editor domain that is an adenosine deaminase variant comprising an alteration selected from the group consisting of R21N, R23H, E25F, N38G, L51W, P54C, M70V, Q71M, N72K, Y73S, M94V, P124W, T133K, D139L, D139M, C146R, and A158K of SEQ ID NO: 1, or a corresponding alteration in another adenosine deaminase.

15 . (canceled)

16 . A fusion protein comprising a polynucleotide programmable DNA binding domain and at least one base editor domain that is an adenosine deaminase variant comprising an alteration V82T and one or more alterations selected from the group consisting of R21N, R23H, E25F, N38G, L51W, P54C, M70V, Q71M, N72K, Y73S, M94V, P124W, T133K, D139L, D139M, C146R, and A158K of SEQ ID NO: 1, or a corresponding alteration in another adenosine deaminase.

17 - 20 . (canceled)

21 . The fusion protein of claim 16 , wherein the adenosine deaminase variant comprises any one of the following groups of alterations:

E25F+V82S+Y123H;

T133K+Y147R+Q154R;

E25F+V82S+Y123H+Y147R+Q154R;

L51W+V82S+Y123H+C146R+Y147R+Q154R;

Y73S+V82S+Y123H+Y147R+Q154R;

P54C+V82S+Y123H+Y147R+Q154R;

N38G+V82T+Y123H+Y147R+Q154R;

N72K+V82S+Y123H+D139L+Y147R+Q154R;

E25F+V82S+Y123H+D139M+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

E25F+V82S+Y123H+T133K+Y147R+Q154R;

E25F+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+P124W+Y147R+Q154R;

L51W+V82S+Y123H+C146R+Y147R+Q154R;

P54C+V82S+Y123H+Y147R+Q154R;

Y73S+V82S+Y123H+Y147R+Q154R;

N38G+V82T+Y123H+Y147R+Q154R;

R23H+V82S+Y123H+Y147R+Q154R;

R21N+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+Y147R+Q154R+A158K;

N72K+V82S+Y123H+D139L+Y147R+Q154R;

E25F+V82S+Y123H+D139M+Y147R+Q154R;

M70V+V82S+M94V+Y123H+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

E25F+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82T+Y123H+Y147R+Q154R;

N38G+I76Y+V82S+Y123H+Y147R+Q154R;

R23H+I76Y+V82S+Y123H+Y147R+Q154R;

P54C+I76Y+V82S+Y123H+Y147R+Q154R;

R21N+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82S+Y123H+D139M+Y147R+Q154R;

Y73S+I76Y+V82S+Y123H+Y147R+Q154R;

E25F+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82T+Y123H+Y147R+Q154R;

N38G+I76Y+V82S+Y123H+Y147R+Q154R;

R23H+I76Y+V82S+Y123H+Y147R+Q154R;

P54C+I76Y+V82S+Y123H+Y147R+Q154R;

R21N+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82S+Y123H+D139M+Y147R+Q154R;

Y73S+I76Y+V82S+Y123H+Y147R+Q154R;

V82S+Q154R;

N72K+V82S+Y123H+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R+A158K;

M70V+Q71M+N72K+V82S+Y123H+Y147R+Q154R;

N72K+V82S+Y123H+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

M70V+V82S+M94V+Y123H+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R+A158K; or

M70V+Q71M+N72K+V82S+Y123H+Y147R+Q154R.

22 - 31 . (canceled)

32 . A fusion protein comprising a polynucleotide programmable DNA binding domain comprising the following sequence:

EIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVR

KVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFMQPTV

AYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII

KLPKYSLFELENGRKRMLASAKFLQKGNELALPSKYVNFLYLASHYEKLKGSPED

NEQKQLFVEQHKHYLDEHIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAEN

IIHLFTLTNLGAPRAFKYFDTTIARKEYRSTKEVLDATLIHQSITGLYETRIDLSQLG

GD GGSGGSGGSGGSGGSGGSGGM DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVL

GNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMA

KVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTD

KADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS

GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAED

AKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLS

ASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFY

KFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFY

PFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGAS

AQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGE

QKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKI

IKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRY

TGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQ

VSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQ

TTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMY

VDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKM

KNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI

LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN

AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQ EGADKRTADGSEFESPKKK

RKV *

wherein the bold sequence indicates sequence derived from Cas9, the italics sequence denotes a linker sequence, and the underlined sequence denotes a bipartite nuclear localization sequence, and at least one base editor domain comprising an adenosine deaminase variant comprising an alteration at an amino acid position selected from the group consisting of 21, 23, 25, 38, 51, 54, 70, 71, 72, 73, 94, 124, 133, 138, 139, 146, and 158 of SEQ ID NO: 1.

33 - 38 . (canceled)

39 . The fusion protein of claim 32 , wherein the adenosine deaminase variant comprises any one of the following groups of alterations:

E25F+V82S+Y123H;

T133K+Y147R+Q154R;

E25F+V82S+Y123H+Y147R+Q154R;

L51W+V82S+Y123H+C146R+Y147R+Q154R;

Y73S+V82S+Y123H+Y147R+Q154R;

P54C+V82S+Y123H+Y147R+Q154R;

N38G+V82T+Y123H+Y147R+Q154R;

N72K+V82S+Y123H+D139L+Y147R+Q154R;

E25F+V82S+Y123H+D139M+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

E25F+V82S+Y123H+T133K+Y147R+Q154R;

E25F+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+P124W+Y147R+Q154R;

L51W+V82S+Y123H+C146R+Y147R+Q154R;

P54C+V82S+Y123H+Y147R+Q154R;

Y73S+V82S+Y123H+Y147R+Q154R;

N38G+V82T+Y123H+Y147R+Q154R;

R23H+V82S+Y123H+Y147R+Q154R;

R21N+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+Y147R+Q154R+A158K;

N72K+V82S+Y123H+D139L+Y147R+Q154R;

E25F+V82S+Y123H+D139M+Y147R+Q154R;

M70V+V82S+M94V+Y123H+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

E25F+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82T+Y123H+Y147R+Q154R;

N38G+I76Y+V82S+Y123H+Y147R+Q154R;

R23H+I76Y+V82S+Y123H+Y147R+Q154R;

P54C+I76Y+V82S+Y123H+Y147R+Q154R;

R21N+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82S+Y123H+D139M+Y147R+Q154R;

Y73S+I76Y+V82S+Y123H+Y147R+Q154R;

E25F+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82T+Y123H+Y147R+Q154R;

N38G+I76Y+V82S+Y123H+Y147R+Q154R;

R23H+I76Y+V82S+Y123H+Y147R+Q154R;

P54C+I76Y+V82S+Y123H+Y147R+Q154R;

R21N+I76Y+V82S+Y123H+Y147R+Q154R;

I76Y+V82S+Y123H+D139M+Y147R+Q154R;

Y73S+I76Y+V82S+Y123H+Y147R+Q154R;

V82S+Q154R;

N72K+V82S+Y123H+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R+A158K;

M70V+Q71M+N72K+V82S+Y123H+Y147R+Q154R;

N72K+V82S+Y123H+Y147R+Q154R;

Q71M+V82S+Y123H+Y147R+Q154R;

M70V+V82S+M94V+Y123H+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R;

V82S+Y123H+T133K+Y147R+Q154R+A158K;

M70V+Q71M+N72K+V82S+Y123H+Y147R+Q154R; or

or any other alteration or group thereof of Table 14.

40 - 60 . (canceled)

61 . A polynucleotide encoding the fusion protein of claim 32 .

62 . A cell produced by introducing into the cell, or a progenitor thereof:

a polynucleotide encoding the fusion protein of claim 12 , and

one or more guide polynucleotides that target the base editor to effect an A•T to GC alteration of a SNP associated with a genetic disease.

63 - 66 . (canceled)

67 . An isolated cell or population of cells propagated or expanded from the cell of claim 62 .

68 . A method of treating a genetic disease in a subject in need thereof, the method comprising administering to the subject a cell of claim 62 .

69 . (canceled)

70 . A base editor system comprising a polynucleotide programmable DNA binding domain and at least one base editor domain that is an adenosine deaminase variant comprising an alteration at an amino acid position selected from the group consisting of 21, 23, 25, 38, 51, 54, 70, 71, 72, 73, 82, 94, 124, 133, 139, 146, and 158 of SEQ ID NO: 1, or a corresponding alteration in another adenosine deaminase:

(SEQ ID NO: 1)

        10         20         30         40

MSEVEFSHEY WMRHALTLAK  R A R D E REVPV GAVLVLN N RV

        50         60         70         80

IGEGWNRAIG  L HD P TAHAEI MALRQGGLV M   QNY RLIDATL

        90        100        110        120

YVTFEPCVMC AGA M IHSRIG RVVFGVRNAK TGAAGSLMDV

       130        140        150        160

LHYPGMNHRV EI T EGILA D E GAALL C YFER MPRQVFN A QK

KAQSSTD.

71 - 86 . (canceled)

87 . A method for correcting a single nucleotide polymorphism (SNP) in a polynucleotide:

contacting a target nucleotide sequence, at least a portion of which is located in the polynucleotide or its reverse complement, with a fusion protein of claim 12 ; and editing the SNP by deaminating the SNP or its complement nucleobase upon targeting of the base editor to the target nucleotide sequence, wherein deaminating the SNP or its complement nucleobase corrects the SNP.

88 - 89 . (canceled)

90 . A method for editing a polynucleotide, the method comprising contacting a target nucleotide sequence with the fusion protein of claim 12 , thereby editing the polynucleotide.

91 - 92 . (canceled)

93 . A base editor comprising an ABE9 comprising a TadA*7.10 adenosine deaminase variant domain and a Cas9 endonuclease domain selected from the following:

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+A109S of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+T111R of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+D119N of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+H122N of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147d+Q154S of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+F149Y of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+T166I of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER); and

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+D167N of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER).

mono TadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+L36H+N157K of SEQ ID NO: 1, and spCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G (MQKFRAER);

mono TadA*7.10 having mutations I76Y+V82T+Y147D+Q154S+F149Y+D167N+L36H+N157K of SEQ ID NO: 1, and SpCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147D+Q154S+F149Y+D167N+L36H+N157K+V106W of SEQ ID NO: 1, and SpCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G (MQKFRAER);

mono TadA*7.10 having mutations A109S+T111R+D119N+H122N+Y147D+F149Y+T166I+D167N of SEQ ID NO: 1, and SpCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G, MQKFRAER; and

mono TadA*7.10 having mutations A109S+T111R+D119N+H122N+Y147D+F149Y+T166I+D167N+V106W of SEQ ID NO: 1, and SpCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G (MQKFRAER); and one or more guide polynucleotides that target the adenosine deaminase variant domain to effect an A•T to G•C alteration of a SNP associated with a genetic disease.

94 . (canceled)

95 . A vector comprising one or more polynucleotides encoding an ABE9 base editor comprising a TadA adenosine deaminase domain and an SpCas9 endonuclease domain selected from monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+A109S and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+T111R and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+D119N and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+H122N and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147d+Q154S and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+F149Y and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+T166I and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER); and

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+D167N and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER).

mono TadA*7.10 having mutations I76Y+V82T+Y147T+Q154S+L36H+N157K and spCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G (MQKFRAER);

mono TadA*7.10 having mutations I76Y+V82T+Y147D+Q154S+F149Y+D167N+L36H+N157K and SpCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T+Y147D+Q154S+F149Y+D167N+L36H+N157K+V106W and SpCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G, (MQKFRAER)

mono TadA*7.10 having mutations A109S+T111R+D119N+H122N+Y147D+F149Y+T166I+D167N and SpCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G (MQKFRAER); and

mono TadA*7.10 having mutations A109S+T111R+D119N+H122N+Y147D+F149Y+T166I+D167N+V106W and SpCas9 having mutations I322V, S409I, E427G, R654L, R753G, R1114G (MQKFRAER).

96 . (canceled)

97 . A composition comprising the fusion protein of claim 12 .

98 . (canceled)

99 . A composition comprising the fusion protein of claim 12 .

100 . A composition comprising the base editor system of claim 70 , wherein the guide RNA comprises a nucleic acid sequence that is complementary to an SERPINA1 gene associated with alpha-1 antitrypsin deficiency (A1AD).

101 - 103 . (canceled)

104 . A pharmaceutical composition for the treatment of a disease or disorder comprising the fusion protein of claim 1 .

105 - 114 . (canceled)

115 . A method of treating alpha-1 antitrypsin deficiency (A1AD), the method comprising administering to a subject in need thereof the pharmaceutical composition of claim 104 .

116 - 119 . (canceled)

120 . An adenosine deaminase variant which is a TadA*7.10 variant comprising any one of the following amino acid alterations or groups of alterations:

V82T;

I76Y+V82T; or

I76Y+V82T+Y147T+Q154S.

121 . A fusion protein comprising a polynucleotide programmable DNA binding domain and at least one base editor domain that is an TadA*7.10 adenosine deaminase variant comprising any one of the following amino acid alterations or groups of alterations:

V82T;

I76Y+V82T; or

I76Y+V82T+Y147T+Q154S.

122 - 124 . (canceled)

125 . A nucleobase editor comprising a TadA*7.10 adenosine deaminase variant domain and a Cas9 endonuclease domain selected from the following:

monoTadA*7.10 having mutation V82T and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER);

monoTadA*7.10 having mutations I76Y+V82T and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER); or

monoTadA*7.10 having mutations I76Y+V82T+Y147T+Q154S and spCas9 having mutations I322V, S409I, E427G, R654L, R753G (MQKFRAER).

Assignments (4)
SECURITY INTEREST Recorded Mar 6, 2026
From: BEAM THERAPEUTICS INC.
To: SIXTH STREET LENDING PARTNERS, AS ADMINISTRATIVE AGENT
Reel/Frame 075021/0929 →
SECURITY INTEREST Recorded Feb 24, 2026
From: BEAM THERAPEUTICS INC.; GUIDE THERAPEUTICS, LLC; BBBR, LLC
To: SIXTH STREET LENDING PARTNERS, AS ADMINISTRATIVE AGENT
Reel/Frame 074955/0064 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 24, 2023
From: GAUDELLI, NICOLE; PACKER, MICHAEL
To: BEAM THERAPEUTICS INC.
Reel/Frame 063091/0128 →
CHANGE OF ADDRESS Recorded Mar 24, 2023
From: BEAM THERAPEUTICS INC.
To: BEAM THERAPEUTICS INC.
Reel/Frame 063163/0245 →