Hi everyone,
I tried to use CRISPR seek to design gRNA for pig gene. Below error occurred. However, there are no txdb for pig genome existing. How to figure it out?
Error in offTargetAnalysis(inputFilePath2, findgRNAsWithREcutOnly = FALSE, :
To indicate whether an offtarget is inside an exon, txdb is
required as TxDb object!
In addition: Warning message:
In dir.create(outputDir) :
'D:\RNA seq analysis R\CRISPRseek\Pig Chrom1 gRNA' already exists
>
results_Pig2 <- offTargetAnalysis(inputFilePath2, findgRNAsWithREcutOnly = FALSE,
REpatternFile = REpatternFile,findPairedgRNAOnly = FALSE,
BSgenomeName = BSgenomeName,chromToSearch= "all",
orgAnn = orgAnn,
max.mismatch = 1, outputDir = outputDir, overwrite = TRUE)
Please follow the script from Hervé or James to create the txdb for the pig genome, then set the parameter txdb and orgAnn in the offtargetAnalysis function call, e.g., txdb = txdb.susScr11.UCSC, orgAnn = org.Ss.egSYMBOL after you run the following script. About the warning message, it should go away if you set overwrite = TRUE in the offtargetAnalysis function call.
Hi Julie,
Thank you very much for your quick response. The problem has been resolved by following the script from James and Hervé. I will try several sequences following the vignettes for the design.
Thanks again,
Xin
Also worth mentioning the
makeTxDbFrom*()
functions from the GenomicFeatures package.For example
makeTxDbFromUCSC()
lets you choose your table/track:See
?makeTxDbFromUCSC
for more information.Or use
makeTxDbFromEnsembl()
if you prefer to fetch the gene models directly from Ensembl:See
?makeTxDbFromEnsembl
for more information.Finally note that AnnotationHub hosts a few EnsDb objects which are functionally equivalent to TxDb objects with some additional capabilities:
See documentation in the ensembldb package for more information.
Best,
H.
Thank you very much, James!