Rosa26-loxP-stop-loxP-YFP mice - 未名空间MITBBS历史存档

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Rosa26-loxP-stop-loxP-YFP mice

Rosa26-loxP-stop-loxP-YFP mice# Biology - 生物学

w*12013-03-21 07:03

1 楼

今天信用卡发Email说Verizon preorder iPhone 6s的单charge了，网上order status
一查发现Expected ship date 从9/25改成9/19了，但是还没有tacking number.
如果按照大家说的overnight shipping, 难道9/25前就能收到了？

a*s2013-03-21 07:03

2 楼

Why do some of my Rosa26-loxP-stop-loxP-YFP mice show YFP+ even without Cre
or anything else?? Does anyone have experience with these mice from Jax?
Thanks!

b*t2013-03-21 07:03

3 楼

No way, overnight shipping就是到了家门口也要等到9/25才投递。从来没买过iPhone
？

s*92013-03-21 07:03

4 楼

Just the mice directly from JAX or breeder from your own colony.
If direct from JAX, I would say it is the compensation issue.

Cre

【在 a*****s 的大作中提到】

: Why do some of my Rosa26-loxP-stop-loxP-YFP mice show YFP+ even without Cre
: or anything else?? Does anyone have experience with these mice from Jax?
: Thanks!

w*12013-03-21 07:03

5 楼

从来没有preorder过，今年LD点名要玫瑰金，才赶这么一浑水。

iPhone

【在 b****t 的大作中提到】

: No way, overnight shipping就是到了家门口也要等到9/25才投递。从来没买过iPhone
: ？

f*02013-03-21 07:03

6 楼

我以前做的Rosa26细胞也是这样.这个东西有自发的翻转或删除.而且表达率还不低,拿
第一代都那样.所以我对科学真得失去了兴致,某些新的发现,其实都是假阳性导致的,可
笑的是一班"科学巨头"发表后,徒子徒生们就不得不从各个方面进行cover.

C*i2013-03-21 07:03

7 楼

lol Rose Gold

m*52013-03-21 07:03

8 楼

这个东西一直有改进，比如后来加入了insulator 来防止transcription read through
, 没你说的那么夸张。

【在 f*********0 的大作中提到】

: 我以前做的Rosa26细胞也是这样.这个东西有自发的翻转或删除.而且表达率还不低,拿
: 第一代都那样.所以我对科学真得失去了兴致,某些新的发现,其实都是假阳性导致的,可
: 笑的是一班"科学巨头"发表后,徒子徒生们就不得不从各个方面进行cover.

s*k2013-03-21 07:03

9 楼

我也给老婆女儿定了玫瑰金。给自己还是定了金色，不能太娘炮。

h*n2013-03-21 07:03

10 楼

this is leak, it happens in almost all LoxP-STOP-LoxP system. The stronger
of the promoter it has, the more leak they might have.
transcription machinery is not like 1 or 0, it has some 0.2 or 0.8 status.

Cre

【在 a*****s 的大作中提到】

: Why do some of my Rosa26-loxP-stop-loxP-YFP mice show YFP+ even without Cre
: or anything else?? Does anyone have experience with these mice from Jax?
: Thanks!

w*12013-03-21 07:03

11 楼

Update 一下，已经有Fedex tracking了,现在东西都出了TN了,我们中西部州明晚或者
周二早肯定就到了。
泥马非要等到周五才送我家吗？那Preorder一点优越感都没了，还不如周五早上上班
pass by Verizon Local 店直接拿一个走还快些。

m*52013-03-21 07:03

12 楼

Promoter本来就是一直开着的，不管是r26还是CAG还是CMV还是whatever being used
in this kind of system
这是transcription read through, 通常是因为loxp stop loxp 中的stop太弱

stronger

【在 h********n 的大作中提到】

: this is leak, it happens in almost all LoxP-STOP-LoxP system. The stronger
: of the promoter it has, the more leak they might have.
: transcription machinery is not like 1 or 0, it has some 0.2 or 0.8 status.
:
: Cre

w*12013-03-21 07:03

13 楼

跟我家搭配一样，LD是玫瑰金6s Plus, 我是金色 6s

【在 s****k 的大作中提到】

: 我也给老婆女儿定了玫瑰金。给自己还是定了金色，不能太娘炮。

s*92013-03-21 07:03

14 楼

My two cents:
How protein can be expressed with a stop code, which cause a frame shift?
For my personal experience, I used Rosa-Stop-YFP mice from JAX and cannot
see any leakage in Cre negative mice in any lymphocyte populations.
It's a transgene and thus it is possible that the insert allele can move and
incorporate into other promote region. In that case, you just need to
refresh your line and get a new breeder from JAX.

stronger

【在 h********n 的大作中提到】

s*p2013-03-21 07:03

15 楼

是的等周五才送
以前也有类似情况

【在 w***1 的大作中提到】

: Update 一下，已经有Fedex tracking了,现在东西都出了TN了,我们中西部州明晚或者
: 周二早肯定就到了。
: 泥马非要等到周五才送我家吗？那Preorder一点优越感都没了，还不如周五早上上班
: pass by Verizon Local 店直接拿一个走还快些。

m*52013-03-21 07:03

16 楼

rosa 是在rosa位点的knock in

and

【在 s****9 的大作中提到】

: My two cents:
: How protein can be expressed with a stop code, which cause a frame shift?
: For my personal experience, I used Rosa-Stop-YFP mice from JAX and cannot
: see any leakage in Cre negative mice in any lymphocyte populations.
: It's a transgene and thus it is possible that the insert allele can move and
: incorporate into other promote region. In that case, you just need to
: refresh your line and get a new breeder from JAX.
:
: stronger

s*k2013-03-21 07:03

17 楼

我的是t-mobile，还没拿到tracking number，但对方保证会在周五deliver.

h*n2013-03-21 07:03

18 楼

再说说我的看法吧.
LoxP-stop-LoxP-geneX system will leak more or less. You did not find
protein expression only means the leakage is too low for your detection.
When a LSL-geneX mouse grows to adult and Cre recombinase is introduced by
adeno or lenti, the "newly" expressed geneX will not be recognized as a
foreign protein by mouse immune system.

and

【在 s****9 的大作中提到】

p*w2013-03-21 07:03

19 楼

t-mobile, 我的order状态已经变成6号以后才能deliver了。哭死……

h*n2013-03-21 07:03

20 楼

I guess stronger promoter will tend to have higher chance of transcription
read through.
But I could be wrong. Correct me with ref.

【在 m******5 的大作中提到】

: Promoter本来就是一直开着的，不管是r26还是CAG还是CMV还是whatever being used
: in this kind of system
: 这是transcription read through, 通常是因为loxp stop loxp 中的stop太弱
:
: stronger

z*e2013-03-21 07:03

21 楼

无论你是哪个途径预定的iPhone，最早都是这周五运到。

D*a2013-03-21 07:03

22 楼

看哪个line吧，leak严重么？有的leak对自己的课题也没什么影响，照实写就是了，也
不会因为这个课题就不能做了。

l*h2013-03-21 07:03

23 楼

我order的两个都要下个月才能到，脑残粉的尤其慢啊

l*12013-03-21 07:03

24 楼

RE LZ
pls refer
PMID 21063464
Liu Y et al.
Tamoxifen-independent recombination in the RIP-CreER mouse. (2010).
PLoS One. 25: e13533.
http://www.ncbi.nlm.nih.gov/pubmed/21063464
>There are at least two possible explanations for
>these discrepancies.
>One possibility is that after many generations
of backcrossing to the C57BL/6 background, the expression of the
randomly integrated RIP-CreER transgene is enhanced, thus
increasing the probability of its nuclear entry, even in tamoxifenuntreated
cells. Another more likely possibility is that the Z/AP
cassette in the Z/AP reporter mouse strain is less prone to
recombination, i.e. it is inserted in an area of the chromatin less
amenable to recombination. Easy access of the CreER to the
target locus may result in both high recombination efficiency and
leakiness of the system as seen in the 3 lines described here. On the
other hand, recombination of a less accessible, heterochromatic
locus could be less efficient, but also less leaky, as in the case of the
Z/AP mouse.
>One way to mitigate such problems is the use of the Pdx1-CreER
>mouse, which was also generated in the Melton lab [14].