US 10,378,007 B2Granted

Methods for modulating TAL specificity

AssigneeCellectis

InventorsAlexandre Juillerat, Philippe Duchateau

Life of this patent

Estimating the term…

Abstract

Methods for improving or modulating targeting specificity of TALE proteins by introducing alternative RVDs into their modular nucleic acid binding domains. Polynucleotides encoding TALE proteins having alternative targeting specificity towards a nucleic acid target sequence. TALE proteins having alternative targeting specificity towards a nucleic acid target sequence and methods of making and using them.

Claims8

Dependent claims

A method for synthesizing a polynucleotide sequence encoding a transcription activator-like effector (TALE) protein, said protein having alternative targeting specificity towards a nucleic acid target sequence containing a nucleic acid base(s) G, wherein said method comprises assembling at least 9 polynucleotide sequences encoding RVDs (repeat variable disresidue), each RVD having specificity to a nucleic acid base in said nucleic acid target, and wherein at least one of said 9 RVDs are selected among the alternative RVD code:

NI or NG to target G, and
wherein the RVD is within an RVD triplet selected from:
NI-HD-NI for ATG;
NI-HD-NI for GCA;
NI-HD-NI for GTA;
NI-HD-NN for GCG;
NI-NN-NI for GAA;
NI-NN-HD for GGC;
NI-NN-NN for GAG;
NI-NN-NN for GGG;
NI-NN-NG for AGG;
NI-NN-NG for GGT;
NI-NG-NI for ATG;
NI-NG-NI for GTA;
NI-NG-HD for AGC;
NI-NG-HD for GAC;
NI-NG-HD for GTA;
NI-NG-HD for GTC;
NI-NG-NN for AGG;
NI-NG-NN for GTA;
NI-NG-NN for GTG;
NI-NG-NG for GTT;
HD-NN-NI for AAG;
HD-NN-NI for AGG;
HD-NN-NI for CGG;
HD-NN-NG for AGG;
HD-NN-NG for CGG;
HD-NG-NI for ATG;
HD-NG-NI for CGA;
HD-NG-NI for CTG;
HD-NG-HD for AGC;
HD-NG-NN for AGG;
HD-NG-NN for CGG;
HD-NG-NG for AGT;
HD-NG-NG for ATG;
HD-NG-NG for CGT;
HD-NG-NG for CTG;
NN-NI-NI for AGA;
NN-NI-NI for GGA;
NN-NI-HD for AGC;
NN-NI-HD for GGC;
NN-NI-NN for GGA;
NN-NI-NG for AGT;
NN-NI-NG for GAG;
NN-NI-NG for GGT;
NN-HD-NI for AAG;
NN-HD-NI for ACG;
NN-HD-NI for GCG;
NN-NN-NI for AAG;
NN-NN-NI for AGG;
NN-NN-NI for GAG;
NN-NN-NI for GGG;
NN-NN-NG for AGG;
NN-NN-NG for GAG;
NN-NN-NG for GGG;
NN-NG-NI for AGA;
NN-NG-NI for ATG;
NN-NG-NI for GTG;
NN-NG-HD for AGC;
NN-NG-NN for AGG;
NN-NG-NN for GGG;
NN-NG-NG for ATG;
NN-NG-NG for GGT;
NN-NG-NG for GTG;
NG-NI-NI for GAA;
NG-NI-NI for TGA;
NG-NI-HD for GAC;
NG-NI-HD for TGC;
NG-NI-NN for GAG;
NG-NI-NN for TGA;
NG-NI-NG for AGT;
NG-NI-NG for GAT;
NG-NI-NG for TAG;
NG-NI-NG for TGT;
NG-HD-NI for GCA;
NG-HD-NI for TCG;
NG-HD-HD for GCC;
NG-HD-NN for GCA;
NG-HD-NN for GCG;
NG-NN-NI for GAA;
NG-NN-NI for GGA;
NG-NN-NI for TGG;
NG-NN-HD for GAC;
NG-NN-HD for GCC;
NG-NN-HD for GGC;
NG-NN-NN for GAG;
NG-NN-NN for GGA;
NG-NN-NN for GGG;
NG-NN-NG for GGT;
NG-NN-NG for TAG;
NG-NN-NG for TGG;
NG-NG-HD for GTC;
NG-NG-HD for TGC;
NG-NG-NN for GTA;
NG-NG-NN for GTG;
NG-NG-NN for TGG;
NI-HD-NI for ATG;
NI-HD-NI for GTA;
NI-NG-HD for GAC;
NG-NI-NG for AGT;
and NG-NN-HD for GCC.

98 more elements

A method for synthesizing a nucleic acid target sequence targetable by a transcription activator-like effector (TALE) protein having a RVD sequence containing at least NG or NI, wherein each base of said target sequence is respectively selected to target said RVDs of said transcription activator-like effector (TALE) protein, and wherein, at least one base in said target sequence is G in order to contact NI or NG, and

wherein the RVD is within an RVD triplet selected from:
NI-HD-NI for ATG;
NI-HD-NI for GCA;
NI-HD-NI for GTA;
NI-HD-NN for GCG;
NI-NN-NI for GAA;
NI-NN-HD for GGC;
NI-NN-NN for GAG;
NI-NN-NN for GGG;
NI-NN-NG for AGG;
NI-NN-NG for GGT;
NI-NG-NI for ATG;
NI-NG-NI for GTA;
NI-NG-HD for AGC;
NI-NG-HD for GAC;
NI-NG-HD for GTA;
NI-NG-HD for GTC;
NI-NG-NN for AGG;
NI-NG-NN for GTA;
NI-NG-NN for GTG;
NI-NG-NG for GTT;
HD-NN-NI for AAG;
HD-NN-NI for AGG;
HD-NN-NI for CGG;
HD-NN-NG for AGG;
HD-NN-NG for CGG;
HD-NG-NI for ATG;
HD-NG-NI for CGA;
HD-NG-NI for CTG;
HD-NG-HD for AGC;
HD-NG-NN for AGG;
HD-NG-NN for CGG;
HD-NG-NG for AGT;
HD-NG-NG for ATG;
HD-NG-NG for CGT;
HD-NG-NG for CTG;
NN-NI-NI for AGA;
NN-NI-NI for GGA;
NN-NI-HD for AGC;
NN-NI-HD for GGC;
NN-NI-NN for GGA;
NN-NI-NG for AGT;
NN-NI-NG for GAG;
NN-NI-NG for GGT;
NN-HD-NI for AAG;
NN-HD-NI for ACG;
NN-HD-NI for GCG;
NN-NN-NI for AAG;
NN-NN-NI for AGG;
NN-NN-NI for GAG;
NN-NN-NI for GGG;
NN-NN-NG for AGG;
NN-NN-NG for GAG;
NN-NN-NG for GGG;
NN-NG-NI for AGA;
NN-NG-NI for ATG;
NN-NG-NI for GTG;
NN-NG-HD for AGC;
NN-NG-NN for AGG;
NN-NG-NN for GGG;
NN-NG-NG for ATG;
NN-NG-NG for GGT;
NN-NG-NG for GTG;
NG-NI-NI for GAA;
NG-NI-NI for TGA;
NG-NI-HD for GAC;
NG-NI-HD for TGC;
NG-NI-NN for GAG;
NG-NI-NN for TGA;
NG-NI-NG for AGT;
NG-NI-NG for GAT;
NG-NI-NG for TAG;
NG-NI-NG for TGT;
NG-HD-NI for GCA;
NG-HD-NI for TCG;
NG-HD-HD for GCC;
NG-HD-NN for GCA;
NG-HD-NN for GCG;
NG-NN-NI for GAA;
NG-NN-NI for GGA;
NG-NN-NI for TGG;
NG-NN-HD for GAC;
NG-NN-HD for GCC;
NG-NN-HD for GGC;
NG-NN-NN for GAG;
NG-NN-NN for GGA;
NG-NN-NN for GGG;
NG-NN-NG for GGT;
NG-NN-NG for TAG;
NG-NN-NG for TGG;
NG-NG-HD for GTC;
NG-NG-HD for TGC;
NG-NG-NN for GTA;
NG-NG-NN for GTG;
NG-NG-NN for TGG;
NI-HD-NI for ATG;
NI-HD-NI for GTA;
NI-NG-HD for GAC;
NG-NI-NG for AGT;
and NG-NN-HD for GCC.

97 more elements

A method for modulating the specificity of a recombinant transcription activator-like effector (TALE) protein with respect to a nucleic acid target sequence that includes a nucleic acid base(s) G, wherein said method comprises introducing or replacing at least one RVD in the repeat domain of said TALE protein by NI or NG to target G, and

wherein the RVD is within an RVD triplet selected from:
NI-HD-NI for ATG;
NI-HD-NI for GCA;
NI-HD-NI for GTA;
NI-HD-NN for GCG;
NI-NN-NI for GAA;
NI-NN-HD for GGC;
NI-NN-NN for GAG;
NI-NN-NN for GGG;
NI-NN-NG for AGG;
NI-NN-NG for GGT;
NI-NG-NI for ATG;
NI-NG-NI for GTA;
NI-NG-HD for AGC;
NI-NG-HD for GAC;
NI-NG-HD for GTA;
NI-NG-HD for GTC;
NI-NG-NN for AGG;
NI-NG-NN for GTA;
NI-NG-NN for GTG;
NI-NG-NG for GTT;
HD-NN-NI for AAG;
HD-NN-NI for AGG;
HD-NN-NI for CGG;
HD-NN-NG for AGG;
HD-NN-NG for CGG;
HD-NG-NI for ATG;
HD-NG-NI for CGA;
HD-NG-NI for CTG;
HD-NG-HD for AGC;
HD-NG-NN for AGG;
HD-NG-NN for CGG;
HD-NG-NG for AGT;
HD-NG-NG for ATG;
HD-NG-NG for CGT;
HD-NG-NG for CTG;
NN-NI-NI for AGA;
NN-NI-NI for GGA;
NN-NI-HD for AGC;
NN-NI-HD for GGC;
NN-NI-NN for GGA;
NN-NI-NG for AGT;
NN-NI-NG for GAG;
NN-NI-NG for GGT;
NN-HD-NI for AAG;
NN-HD-NI for ACG;
NN-HD-NI for GCG;
NN-NN-NI for AAG;
NN-NN-NI for AGG;
NN-NN-NI for GAG;
NN-NN-NI for GGG;
NN-NN-NG for AGG;
NN-NN-NG for GAG;
NN-NN-NG for GGG;
NN-NG-NI for AGA;
NN-NG-NI for ATG;
NN-NG-NI for GTG;
NN-NG-HD for AGC;
NN-NG-NN for AGG;
NN-NG-NN for GGG;
NN-NG-NG for ATG;
NN-NG-NG for GGT;
NN-NG-NG for GTG;
NG-NI-NI for GAA;
NG-NI-NI for TGA;
NG-NI-HD for GAC;
NG-NI-HD for TGC;
NG-NI-NN for GAG;
NG-NI-NN for TGA;
NG-NI-NG for AGT;
NG-NI-NG for GAT;
NG-NI-NG for TAG;
NG-NI-NG for TGT;
NG-HD-NI for GCA;
NG-HD-NI for TCG;
NG-HD-HD for GCC;
NG-HD-NN for GCA;
NG-HD-NN for GCG;
NG-NN-NI for GAA;
NG-NN-NI for GGA;
NG-NN-NI for TGG;
NG-NN-HD for GAC;
NG-NN-HD for GCC;
NG-NN-HD for GGC;
NG-NN-NN for GAG;
NG-NN-NN for GGA;
NG-NN-NN for GGG;
NG-NN-NG for GGT;
NG-NN-NG for TAG;
NG-NN-NG for TGG;
NG-NG-HD for GTC;
NG-NG-HD for TGC;
NG-NG-NN for GTA;
NG-NG-NN for GTG;
NG-NG-NN for TGG;
NI-HD-NI for ATG;
NI-HD-NI for GTA;
NI-NG-HD for GAC;
NG-NI-NG for AGT;
and NG-NN-HD for GCC.

97 more elements

Dependencies are inferred from the claim text.

Facts

Application: 14/424,757Filed 30 Aug 2013 · us provisional application
Priority: 3 Sep 2012
Published: US 2015/0218230 A16 Aug 2015
Granted: 13 Aug 2019
Term ends: Not estimatedThe source records don’t support an estimate.

Classification11 codes

C12N 15/10
Recombinant DNA-technology: processes for the isolation, preparation or purification of DNA or RNA
C07H 21/00
Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate
C07K 14/195
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof: from bacteria
C12N 15/102
Processes for the isolation, preparation or purification of DNA or RNA: mutagenizing nucleic acids

Family

Application61/696,254Us provisional application · filed 3 Sep 2012
US 2015/0218230 A1Pre-grant publication · published 6 Aug 2015
US 10,378,007 B2This patent

From the US filing history. Foreign family members aren’t listed.

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Abstract

NI or NG to target G, and
wherein the RVD is within an RVD triplet selected from:
NI-HD-NI for ATG;
NI-HD-NI for GCA;
NI-HD-NI for GTA;
NI-HD-NN for GCG;
NI-NN-NI for GAA;
NI-NN-HD for GGC;
NI-NN-NN for GAG;
NI-NN-NN for GGG;
NI-NN-NG for AGG;
NI-NN-NG for GGT;
NI-NG-NI for ATG;
NI-NG-NI for GTA;
NI-NG-HD for AGC;
NI-NG-HD for GAC;
NI-NG-HD for GTA;
NI-NG-HD for GTC;
NI-NG-NN for AGG;
NI-NG-NN for GTA;
NI-NG-NN for GTG;
NI-NG-NG for GTT;
HD-NN-NI for AAG;
HD-NN-NI for AGG;
HD-NN-NI for CGG;
HD-NN-NG for AGG;
HD-NN-NG for CGG;
HD-NG-NI for ATG;
HD-NG-NI for CGA;
HD-NG-NI for CTG;
HD-NG-HD for AGC;
HD-NG-NN for AGG;
HD-NG-NN for CGG;
HD-NG-NG for AGT;
HD-NG-NG for ATG;
HD-NG-NG for CGT;
HD-NG-NG for CTG;
NN-NI-NI for AGA;
NN-NI-NI for GGA;
NN-NI-HD for AGC;
NN-NI-HD for GGC;
NN-NI-NN for GGA;
NN-NI-NG for AGT;
NN-NI-NG for GAG;
NN-NI-NG for GGT;
NN-HD-NI for AAG;
NN-HD-NI for ACG;
NN-HD-NI for GCG;
NN-NN-NI for AAG;
NN-NN-NI for AGG;
NN-NN-NI for GAG;
NN-NN-NI for GGG;
NN-NN-NG for AGG;
NN-NN-NG for GAG;
NN-NN-NG for GGG;
NN-NG-NI for AGA;
NN-NG-NI for ATG;
NN-NG-NI for GTG;
NN-NG-HD for AGC;
NN-NG-NN for AGG;
NN-NG-NN for GGG;
NN-NG-NG for ATG;
NN-NG-NG for GGT;
NN-NG-NG for GTG;
NG-NI-NI for GAA;
NG-NI-NI for TGA;
NG-NI-HD for GAC;
NG-NI-HD for TGC;
NG-NI-NN for GAG;
NG-NI-NN for TGA;
NG-NI-NG for AGT;
NG-NI-NG for GAT;
NG-NI-NG for TAG;
NG-NI-NG for TGT;
NG-HD-NI for GCA;
NG-HD-NI for TCG;
NG-HD-HD for GCC;
NG-HD-NN for GCA;
NG-HD-NN for GCG;
NG-NN-NI for GAA;
NG-NN-NI for GGA;
NG-NN-NI for TGG;
NG-NN-HD for GAC;
NG-NN-HD for GCC;
NG-NN-HD for GGC;
NG-NN-NN for GAG;
NG-NN-NN for GGA;
NG-NN-NN for GGG;
NG-NN-NG for GGT;
NG-NN-NG for TAG;
NG-NN-NG for TGG;
NG-NG-HD for GTC;
NG-NG-HD for TGC;
NG-NG-NN for GTA;
NG-NG-NN for GTG;
NG-NG-NN for TGG;
NI-HD-NI for ATG;
NI-HD-NI for GTA;
NI-NG-HD for GAC;
NG-NI-NG for AGT;
and NG-NN-HD for GCC.

98 more elements

Facts

Application

14/424,757Filed 30 Aug 2013 · us provisional application

Priority

3 Sep 2012

Published

US 2015/0218230 A16 Aug 2015

Granted

13 Aug 2019

Term ends

Not estimatedThe source records don’t support an estimate.

Classification11 codes

C12N 15/10
Recombinant DNA-technology: processes for the isolation, preparation or purification of DNA or RNA
C07H 21/00
Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate
C07K 14/195
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof: from bacteria
C12N 15/102
Processes for the isolation, preparation or purification of DNA or RNA: mutagenizing nucleic acids