README.md

Generating lwd1 mutants

Using CRISPR-Cas9 to generate lwd1 mutants to confirm LWD1 as the gene underlying the eam7 locus. GP-fast embryos were transformed with vectors carrying sgRNAs that target the start or the end of the LWD1 coding sequence. The resulting T0 plants were genotyped to screen for different induced mutations. Promising T1 plants were sown, and genotyped again. From this, three different homozygous mutants were selected for gene expression analysis and phenotyping: lwd1-26, lwd1-390 and lwd1-402.

Study overview:

   flowchart TD;
	A[GP-fast plants] --> |embryo_transformation | B[T0 mutant plants]
	B --> | DNA_extraction| C[T0 mutant DNA samples]
	C --> | plant_genotyping | D[T0 mutant genotype info]
	D -.-> E
	B --> | plant propagation | E[T1 mutant plants]
	E --> | DNA_extraction | F[T1 mutant DNA samples]
	F --> | plant_genotyping | G[T1 mutant genotype info]
	G -.-> H
	E --> | plant propagation | H[T2 mutant plants]
	H -.-> I[used in studies:
	- gene_expresssion_lwd1_mutants
	- phenotyping_lwd1_mutants]