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TALE library expression vector as well as preparation method and application thereof

An expression vector, TALE-VP64 technology, applied in the field of gene editing, can solve the problem of time-consuming and laborious, achieve the effect of wide application value and time saving

Pending Publication Date: 2022-07-08
SHENYANG UNIV
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  • Application Information

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Problems solved by technology

In this case, more TALEs need to be constructed, which is time-consuming and labor-intensive

Method used

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  • TALE library expression vector as well as preparation method and application thereof
  • TALE library expression vector as well as preparation method and application thereof
  • TALE library expression vector as well as preparation method and application thereof

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Embodiment Construction

[0027] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the embodiments. The specific embodiments described herein are only used to explain the present invention, but not to limit the invention.

[0028] Since members of the TALE family contain a highly conserved and repetitive region, their DNA-binding capacity depends on repeating the 12th and 13th amino acid positions of each module (ie, repeating variable diresidues or RVDs). Within each RVD is a simple code specifying the recognition of a specific base, which determines the specificity of TALE nucleotide binding. Specifically, as figure 1 Shown is the [TALE assembly kit "Golden Gate TALEN and TAL Effector Kit 2.0 (Addgene)" (referred to as Addgene Kit 2.0) for genome engineering.

[0029] (A) Native structures of TALEs derived from Xanthomonas. Each DNA-binding module consists of 34 ami...

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Abstract

The invention discloses a TALE library expression vector as well as a preparation method and application thereof, a PFUS-A + 10 * NH plasmid mutant, a PFUS-B7 + 7 * NH plasmid mutant, a pLR-NH plasmid mutant and a TALE-VP64 skeleton plasmid are assembled into the TALE library expression vector, the assembly is rapid and modularized, the generated RVD content and the whole DNA binding domain composition can be accurately controlled, and the TALE library expression vector has the advantages of high specificity, high specificity, high specificity, high specificity and high efficiency. The TALE library expression vector can be combined with any 18bp target DNA, and the TALE library expression vector disclosed by the invention has a wide application value in the technical fields of gene editing and biology; and researchers can directly use the material as an experimental material, so that a large amount of time is saved for the majority of TALE researchers.

Description

technical field [0001] The invention relates to the field of gene editing, in particular to a TALE library expression vector and a preparation method and application thereof. Background technique [0002] Transcription activator-like effectors (TALEs) are proteins secreted by Xanthomonas that target specific DNA sequences. Members of the TALE family contain a highly conserved and repetitive region, and their DNA-binding capacity depends on repeating the 12th and 13th amino acid positions of each module (ie, repeating variable diresidues or RVDs). Within each RVD is a simple code specifying the recognition of a specific base, which determines the specificity of TALE nucleotide binding. For example, NI(Asn / Ile) corresponds to A, NG(Asn / Gly) corresponds to T, HD(His / Asp) corresponds to C, and NH(Asn / His) corresponds to G. Artificially constructed TALEs that complement DNA-binding domains (DBDs) with functional domains can be applied to many aspects of genetic engineering, inc...

Claims

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Application Information

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IPC IPC(8): C12N15/66C12N15/31C40B50/06C40B40/10
CPCC12N15/66C07K14/195C40B50/06C40B40/10Y02A50/30
Inventor 张书衍王晓微肖延铭蔡思杰郭显阳詹硕
Owner SHENYANG UNIV
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