Site-directed integration of transgenes into the nuclear genome of plants using CRISPR/Cpf1/ssDNA

 

We aim to establish a CRISPR/Cpf1-based technique to insert transgenes into specific genome sites that are predicted to promote high expression.

The Idea

There is great potential in using algae, including the green alga Chlamydomonas reinhardtii, for the sustainable production of high value compounds (Brodie et al., 2017). However, one obstacle in reaching that long-term goal is the limited number of gene editing and genomic transformation technologies for algae. For instance, transformation of the alga Chlamydomonas reinhardtii is via random integration and so requires extensive screening steps to isolate cell lines that stably express the transgene to high levels. To accelerate this process, we aim to establish a CRISPR/Cpf1-based technique (Ferenczi et al., 2017) to insert transgenes into specific genome sites that are predicted to promote high expression. The technique would reduce the variation of gene expression within the transformed cell population, allow high throughput testing of a large number of DNA constructs, and therefore increase our capacity to create highly productive cell lines.

The Team

Nandor D Hegyi,
Summer student, Plant Metabolism Group, Department of Plant Sciences, Cambridge University

Gonzalo I Mendoza-Ochoa,
Plant Metabolism Group, Department of Plant Sciences, Cambridge University

Aleix Gorchs,
Plant Metabolism Group, Department of Plant Sciences, Cambridge University

Payam Mehrshahi,
Plant Metabolism Group, Department of Plant Sciences, Cambridge University

Oleg Raitskin,
Nicola Patron Group, Earlham Institute, Norwich Research Park

Quentin Dudley,
Nicola Patron Group, Earlham Institute, Norwich Research Park


Project Outputs

Project Report

Summary of the project's achievements and future plans

Project Proposal

Original proposal and application

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Project Resources


Jim Haseloff