Mammalian mutagenesis?

March 28, 2006 at 4:37 pm #4212

Participant

Does anyone know if any literature that have effectively isolated a gene in MAMMALIAN cells (via. mutagensis and genetic screening)?

(I know how to do it in drosph, yeast, c. elegans etc etc etc … )

March 28, 2006 at 11:16 pm #44485

canalon

Participant

What do you mean isolated a gene in mammalian cell?

Because if:
– it is taking a gene from a mammal and cloning it, it has been done (recombinant insulin for example).

– it is inserting a gene in a mmal cell, it has also been done (gfp pigs for example, but also goats producing drugs in their milk etc)

otherwise, be a little more precise, but the answer will probably be yes, and you could probably find it on pubmed (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi)

March 30, 2006 at 2:01 am #44607

LilKim

Participant

oK… lemme re-word my question.

I need to set up a genetic screen using mammalian cells. Essentially, I want to use a genetic approach to determine if the RITS (RNAi induced transcriptional silencing) complex exists in mammalian cells (last year it was Identified in yeast and published in Sciencemag)

I did a quick pubmed search looking for any genetic screens performed on mammalian cells. And I essentially came up with nothing (…from respectable journals)

I talked to my professor, and he says it’s do-able… i’ll just have to figure out how the hell to do it??? (this is actually for an assignment.. i have to write a proposal)

… it’s seems like it would be really difficult technically… (i’m already seeing sleepless nights… trying to design this) So, I was wondering if anyone had more of a clue than I do… and if they could provide a clue for me to start from.

thanks in advance:
– KIM

April 10, 2006 at 4:25 pm #45589

Cloak & Dagger

Participant

You could look up the DNA sequence of the yeast components of the complex and search for homologues in a mammalian genome.

Then you could target whatever you find for mutagenesis and screen for RNAi induced silencing. That should confirm whether they have the same functions in mammals (at least in the type of cells you are using).

Hope that helps.

April 10, 2006 at 6:17 pm #45593

LilKim

Participant

Hey.. thanks for the reply

…. my paper is due in another 2 weeks… and I actually thought of really similar idea.

yeah, i’ve actually gone and looked for homologs and orthologs and they haven’t been identified in mammals. BUT, based on the stuff i’ve read .. i think it’s VERY plausible for the RITS activity in mammalian cells…

I’ve devised a plan (along the same-lines as what you’ve said) to solve the problem, and I think it will work. (i’m going to make an appointment to speak with my professor about it..on thursday or friday)

… but, thanks! It’s a good feeling to know that 2 different people came to similar conclusions independantly… so I feel encouraged

thanks again & best wishes!
– KIM

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