Transgenic Monkeys Created Using CRISPR Technology

Why Is CRISPR An Important Breakthrough for Creating Transgenic Monkeys?

Most traditional methods for gene targeting require the introduction of a transgene cassette via homologous recombination. Recombination occurs rarely, so these methods typically use many individual animals – which is challenging to accomplish with non-human primates for both ethical and logistical/budgetary reasons. Successful recombination must be followed by selective breeding, which is very slow in primates compared to other model organisms; monkeys take years to reach reproductive age – compared to only weeks in mice or days in flies.

Viral vectors allow highly efficient transgene delivery, and have been used successfully in primates – including human patients receiving gene therapy! However, it is difficult with viral vectors to control the number of copies inserted into the genome and the location of those gene insertions – which may be okay when you just want to express a therapeutic gene in a specific cell type in an adult individual, but creates challenges when you want to make a heritable change to the genome in order to propagate a transgenic strain of laboratory animals for reproducible long-term experimental use. Without precise control over the insertion site, virally-mediated gene delivery can have unintended consequences by altering the expression of off-target genes whose coding or regulatory sequences get interrupted, and copy-number variation can create major challenges in interpreting experimental results.

CRISPR allows precise targeting of gene mutations – including deletions, alterations, or insertions of novel DNA sequences and is much faster and simpler than earlier techniques such as TALEN, Zinc-Finger nuclease, and Cre-Lox-mediated recombination. Because they use synthetic guide RNA complementary to the targeted sequence, CRISPR-associated nucleases like Cas9 have a much lower rate of off-target effects than viral vectors.

Check Out These Recent Papers That Used CRISPR/Cas9 in Several Different Species

How Does It Work?

The CRISPR method relies upon customized gene constructs that encode a codon-optimized Cas9 nuclease and a synthetic guide RNA for precise targeting.

GenScript's gene synthesis service can prepare the constructs you need and deliver them in as few as 4 business days.

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